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The chemical structure impairs the intensity of genotoxic effects promoted by 1,2-unsaturated pyrrolizidine alkaloids in vitro. Food Chem Toxicol 2022; 164:113049. [PMID: 35500694 DOI: 10.1016/j.fct.2022.113049] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2022] [Revised: 04/05/2022] [Accepted: 04/15/2022] [Indexed: 11/23/2022]
Abstract
1,2-unsaturated pyrrolizidine alkaloids (PAs) represent a large group of secondary plant metabolites exhibiting hepatotoxic, genotoxic, and carcinogenic properties upon bioactivation. To examine how the degree of esterification affects the genotoxic profile of PA we investigated cytotoxicity, histone H2AX phosphorylation, DNA strand break induction, cell cycle perturbation, micronuclei formation, and aneugenic effects in different cell models. Analysis of cytotoxicity and phosphorylation of histone H2AX was structure- and concentration-dependent: diester-type PAs (except monocrotaline) showed more pronounced effects than monoester-type PAs. Cell cycle analysis identified that diester-type PAs induced a S-phase arrest and a decrease in the occurrence of cells in the G1-phase. The same structure-dependency was observed by flow-cytometric analysis of PA-induced micronuclei in CYP3A4-overexpressing V79 cells. Analysis of centromeres induced by lasiocarpine in the micronuclei by fluorescence in situ hybridization indicated an aneugenic effect in V79h3A4 cells. Comet assays revealed no significant induction of DNA strand breaks for all investigated PAs. Overall, diester-type PAs induced more pronounced effects than monoester-type PAs. Furthermore, our results indicate aneugenic effects upon exposure towards lasiocarpine in vitro. These data improve our understanding how structural features of PA influence the genotoxic profile. Especially, the monoester-type PAs seem to induce less severe effects than other PAs.
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Takeishi K, Collin de l'Hortet A, Wang Y, Handa K, Guzman-Lepe J, Matsubara K, Morita K, Jang S, Haep N, Florentino RM, Yuan F, Fukumitsu K, Tobita K, Sun W, Franks J, Delgado ER, Shapiro EM, Fraunhoffer NA, Duncan AW, Yagi H, Mashimo T, Fox IJ, Soto-Gutierrez A. Assembly and Function of a Bioengineered Human Liver for Transplantation Generated Solely from Induced Pluripotent Stem Cells. Cell Rep 2021; 31:107711. [PMID: 32492423 DOI: 10.1016/j.celrep.2020.107711] [Citation(s) in RCA: 85] [Impact Index Per Article: 21.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/05/2019] [Revised: 12/17/2019] [Accepted: 05/08/2020] [Indexed: 12/22/2022] Open
Abstract
The availability of an autologous transplantable auxiliary liver would dramatically affect the treatment of liver disease. Assembly and function in vivo of a bioengineered human liver derived from induced pluripotent stem cells (iPSCs) has not been previously described. By improving methods for liver decellularization, recellularization, and differentiation of different liver cellular lineages of human iPSCs in an organ-like environment, we generated functional engineered human mini livers and performed transplantation in a rat model. Whereas previous studies recellularized liver scaffolds largely with rodent hepatocytes, we repopulated not only the parenchyma with human iPSC-hepatocytes but also the vascular system with human iPS-endothelial cells, and the bile duct network with human iPSC-biliary epithelial cells. The regenerated human iPSC-derived mini liver containing multiple cell types was tested in vivo and remained functional for 4 days after auxiliary liver transplantation in immunocompromised, engineered (IL2rg-/-) rats.
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Affiliation(s)
- Kazuki Takeishi
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan
| | | | - Yang Wang
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Hepatobiliary Surgery, Peking University People's Hospital, Beijing 100044, China
| | - Kan Handa
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Jorge Guzman-Lepe
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Kentaro Matsubara
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Kazutoyo Morita
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Sae Jang
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Nils Haep
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Rodrigo M Florentino
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Physiology and Biophysics, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil
| | - Fangchao Yuan
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China
| | - Ken Fukumitsu
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Kimimasa Tobita
- Department of Bioengineering and Department of Developmental Biology, University of Pittsburgh, Pittsburgh, PA 15201, USA
| | - Wendell Sun
- LifeCell Corporation, Branchburg, NJ 08876, USA
| | - Jonathan Franks
- Center for Biologic Imaging, University of Pittsburgh Medical School, Pittsburgh, PA 15261, USA
| | - Evan R Delgado
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Erik M Shapiro
- Department of Radiology, Michigan State University, East Lansing, MI 48824, USA
| | - Nicolas A Fraunhoffer
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Facultad de Ciencias de la Salud, Carrera de Medicina, Universidad Maimónides, Ciudad Autónoma de Buenos Aires and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Ciudad Autónoma de Buenos Aires, Buenos Aires 1001, Argentina
| | - Andrew W Duncan
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA
| | - Hiroshi Yagi
- Department of Surgery, School of Medicine, Keio University, Tokyo 160-8582, Japan
| | - Tomoji Mashimo
- Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, University of Tokyo, Tokyo 158-8557, Japan
| | - Ira J Fox
- McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Surgery, Children's Hospital of Pittsburgh of UPMC, University of Pittsburgh, Pittsburgh, PA 15224, USA
| | - Alejandro Soto-Gutierrez
- Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA.
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Marongiu F, Laconi E. Cell competition in liver carcinogenesis. World J Hepatol 2020; 12:475-484. [PMID: 32952874 PMCID: PMC7475782 DOI: 10.4254/wjh.v12.i8.475] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/03/2020] [Revised: 06/22/2020] [Accepted: 07/26/2020] [Indexed: 02/06/2023] Open
Abstract
Cell competition is now a well-established quality control strategy to optimize cell and tissue fitness in multicellular organisms. While pursuing this goal, it is also effective in selecting against altered/defective cells with putative (pre)-neoplastic potential, thereby edging the risk of cancer development. The flip side of the coin is that the molecular machinery driving cell competition can also be co-opted by neoplastic cell populations to expand unchecked, outside the boundaries of tissue homeostatic control. This review will focus on information that begins to emerge regarding the role of cell competition in liver physiology and pathology. Liver repopulation by normal transplanted hepatocytes is an interesting field of investigation in this regard. The biological coordinates of this process share many features suggesting that cell competition is a driving force for the clearance of endogenous damaged hepatocytes by normal donor-derived cells, as previously proposed. Intriguing analogies between liver repopulation and carcinogenesis will be briefly discussed and the potential dual role of cell competition, as a barrier or a spur to neoplastic development, will be considered. Cell competition is in essence a cooperative strategy organized at tissue level. One facet of such cooperative attitude is expressed in the elimination of altered cells which may represent a threat to the organismal community. On the other hand, the society of cells can be disrupted by the emergence of selfish clones, exploiting the molecular bar codes of cell competition, thereby paving their way to uncontrolled growth.
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Affiliation(s)
- Fabio Marongiu
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari 09124, Italy
| | - Ezio Laconi
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari 09124, Italy
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Aydın O, Pehlivanlı F, Karaca G, Aydın G, Altunkaya C, Bulut H. May dexpanthenol, platelet-rich plasma, and thymoquinone provide new hope to maintain liver regeneration after partial hepatectomy? TURKISH JOURNAL OF GASTROENTEROLOGY 2020; 30:826-834. [PMID: 31530526 DOI: 10.5152/tjg.2019.18697] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/25/2022]
Abstract
BACKGROUND/AIMS Complete liver regeneration may not always be possible after liver injuries and/or partial liver resection. The present study investigated the effects of dexpanthenol, platelet-rich plasma (PRP), and thymoquinone on liver regeneration in rats after partial hepatectomy (PH). MATERIALS AND METHODS A total of 34 Wistar albino rats, each weighing 250-280 g, were randomly separated into four groups. PH was performed, and except for the control group, intraperitoneal dexpanthenol, PRP, or thymoquinone was administered to the relevant groups for 7 days. All rats were then sacrificed, and the liver tissues were examined histopathologically and biochemically. RESULTS PRP reduced all oxidant-antioxidant parameters in rats that experienced liver regeneration, but did not create histopathological improvement in the liver tissue. Dexpanthenol had a histopathological improving effect on the liver tissue, but had no effect on biochemical parameters. Thymoquinone showed no histopathological or biochemical effects on liver regeneration. CONCLUSION Although dexpanthenol did not affect biochemical oxidative parameters, it was considered to have improving effects on liver regeneration histopathologically. In addition, it was thought that PRP may be used for treatment of ischemia-reperfusion injury and cholestatic damage of the liver. Nevertheless, further studies are required on these subjects.
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Affiliation(s)
- Okan Aydın
- Department of General Surgery, Kırıkkale University School of Medicine, Kırıkkale, Turkey
| | - Faruk Pehlivanlı
- Department of General Surgery, Kırıkkale University School of Medicine, Kırıkkale, Turkey
| | - Gökhan Karaca
- Department of General Surgery, Kırıkkale University School of Medicine, Kırıkkale, Turkey
| | - Gülçin Aydın
- Department of Anesthesiology and Reanimation, Kırıkkale University School of Medicine, Kırıkkale, Turkey
| | - Canan Altunkaya
- Department of Pathology, Kırıkkale University School of Medicine, Kırıkkale, Turkey
| | - Huri Bulut
- Department of Medical Biochemistry, Bezmialem Vakıf University, İstanbul, Turkey
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Czekaj P, Król M, Limanówka Ł, Michalik M, Lorek K, Gramignoli R. Assessment of animal experimental models of toxic liver injury in the context of their potential application as preclinical models for cell therapy. Eur J Pharmacol 2019; 861:172597. [PMID: 31408648 DOI: 10.1016/j.ejphar.2019.172597] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/02/2019] [Revised: 08/04/2019] [Accepted: 08/08/2019] [Indexed: 02/06/2023]
Abstract
Preclinical animal models allow to study development and progression of several diseases, including liver disorders. These studies, for ethical reasons and medical limits, are impossible to carry out in human patients. At the same time, such experimental models constitute an important source of knowledge on pathomechanisms for drug- and virus-induced hepatotoxicity, both acute and chronic. Carbon tetrachloride, D-Galactosamine, and retrorsine are xenobiotics that can be used in immunocompetent animal models of hepatotoxicity, where chemical-intoxicated livers present histological features representative of human viruses-related infection. A prolonged derangement into liver architecture and functions commonly lead to cirrhosis, eventually resulting in hepatocellular carcinoma. In human, orthotopic liver transplantation commonly resolve most the problems related to cirrhosis. However, the shortage of donors does not allow all the patients in the waiting list to receive an organ on time. A promising alternative treatment for acute and chronic liver disease has been advised in liver cell transplantation, but the limited availability of hepatocytes for clinical approaches, in addition to the immunosuppressant regiment required to sustain cellular long-term engraftment have been encouraging the use of alternative cell sources. A recent effective source of stem cells have been recently identified in the human amnion membrane. Human amnion epithelial cells (hAEC) have been preclinically tested and proven sufficient to rescue immunocompetent rodents lethally intoxicated with drugs. The adoption of therapeutic procedures based on hAEC transplant in immunocompetent recipients affected by liver diseases, as well as patients with immune-related disorders, may constitute a successful new alternative therapy in regenerative medicine.
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Affiliation(s)
- Piotr Czekaj
- Department of Cytophysiology, Chair of Histology and Embryology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland, Medyków 18 str., 40-752, Katowice, Poland.
| | - Mateusz Król
- Students Scientific Society, Department of Cytophysiology, Chair of Histology and Embryology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland, Medyków 18 str., 40-752, Katowice, Poland.
| | - Łukasz Limanówka
- Students Scientific Society, Department of Cytophysiology, Chair of Histology and Embryology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland, Medyków 18 str., 40-752, Katowice, Poland
| | - Marcin Michalik
- Students Scientific Society, Department of Cytophysiology, Chair of Histology and Embryology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland, Medyków 18 str., 40-752, Katowice, Poland
| | - Katarzyna Lorek
- Students Scientific Society, Department of Cytophysiology, Chair of Histology and Embryology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland, Medyków 18 str., 40-752, Katowice, Poland
| | - Roberto Gramignoli
- Department of Laboratory Medicine (LABMED), H5, Division of Pathology, Karolinska Institutet, Alfred Nobels Allé 8, 14152, Huddinge, Sweden.
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Clinical hepatocyte transplantation. GASTROENTEROLOGIA Y HEPATOLOGIA 2019; 42:202-208. [DOI: 10.1016/j.gastrohep.2018.10.007] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/23/2018] [Revised: 09/21/2018] [Accepted: 10/10/2018] [Indexed: 12/18/2022]
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The fate of hepatocyte cell line derived from a liver injury model with long-term in vitro passage. Mol Cell Toxicol 2018. [DOI: 10.1007/s13273-018-0029-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/28/2022]
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Marongiu F, Serra MP, Fanti M, Cadoni E, Serra M, Laconi E. Regenerative Medicine: Shedding Light on the Link between Aging and Cancer. Cell Transplant 2017; 26:1530-1537. [PMID: 29113461 PMCID: PMC5680953 DOI: 10.1177/0963689717721224] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/29/2016] [Revised: 02/17/2017] [Accepted: 02/22/2017] [Indexed: 01/07/2023] Open
Abstract
The evidence linking aging and cancer is overwhelming. Findings emerging from the field of regenerative medicine reinforce the notion that aging and cancer are profoundly interrelated in their pathogenetic pathways. We discuss evidence to indicate that age-associated alterations in the tissue microenvironment contribute to the emergence of a neoplastic-prone tissue landscape, which is able to support the selective growth of preneoplastic cell populations. Interestingly, tissue contexts that are able to select for the growth of preneoplastic cells, including the aged liver microenvironment, are also supportive for the clonal expansion of normal, homotypic, transplanted cells. This suggests that the growth of normal and preneoplastic cells is possibly driven by similar mechanisms, implying that strategies based on principles of regenerative medicine might be applicable to modulate neoplastic disease.
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Affiliation(s)
- Fabio Marongiu
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
| | - Maria Paola Serra
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
| | - Maura Fanti
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
| | - Erika Cadoni
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
| | - Monica Serra
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
| | - Ezio Laconi
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
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Rohn S, Schroeder J, Riedel H, Polenz D, Stanko K, Reutzel-Selke A, Tang P, Brusendorf L, Raschzok N, Neuhaus P, Pratschke J, Sawitzki B, Sauer IM, Mogl MT. Allogeneic Liver Transplantation and Subsequent Syngeneic Hepatocyte Transplantation in a Rat Model: Proof of Concept for in vivo Tissue Engineering. Cells Tissues Organs 2016; 201:399-411. [DOI: 10.1159/000445792] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 03/25/2016] [Indexed: 11/19/2022] Open
Abstract
Objectives: Stable long-term functioning of liver cells after transplantation in humans is still not achieved successfully. A new approach for successful engraftment of liver cells may be the transplantation of syngeneic cells into an allogeneic liver graft. We therefore developed a new rat model for combined liver and liver cell transplantation (cLCTx) under stable immunosuppression. Materials and Methods: After inducing a mitotic block, liver grafts from female donor rats (Dark Agouti) were transplanted into female recipients (Lewis). In male Lewis rats, liver cell proliferation was induced with subsequent cell isolation and transplantation into female recipients after organ transplantation. Y-chromosome detection of the transplanted male cells was performed by quantitative polymerase chain reaction (qPCR) and fluorescence in situ hybridization (FisH) with localization of transplanted cells by immunohistochemistry. Results: Immunohistochemistry demonstrated the engraftment of transplanted cells, as confirmed by FisH, showing repopulation of the liver graft with 15.6% male cells (± 1.8 SEM) at day 90. qPCR revealed 14.15% (± 5.09 SEM) male DNA at day 90. Conclusion: Engraftment of transplanted syngeneic cells after cLCTx was achieved for up to 90 days under immunosuppression. Immunohistochemistry indicated cell proliferation, and the FisH results were partly confirmed by qPCR. This new protocol in rats appears feasible for addressing long-term functioning and eventually the induction of operational tolerance in the future.
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Effect of nebivolol on liver regeneration in an experimental 70% partial hepatectomy model. Asian J Surg 2016; 40:375-379. [PMID: 26920216 DOI: 10.1016/j.asjsur.2015.12.005] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/14/2015] [Revised: 12/22/2015] [Accepted: 12/30/2015] [Indexed: 12/16/2022] Open
Abstract
BACKGROUND Factors affecting liver regeneration are still relevant. The purpose of this study is to investigate the effect of nebivolol treatment on liver regeneration in rats in which 70% partial hepatectomy was performed. METHODS Three groups were created: the control group, the low dose group, and the high dose group, with 20 rats in each group and 70% hepatectomy was performed in all rats. Immediately after partial liver resection, 2 mL physiological saline solution was administered to the control group via oral gavage, 0.5 mg/kg nebivolol was administered via oral gavage to the low dose group and 2 mg/kg nebivolol was administered via oral gavage to the high dose group. On the 1st and 5th days after liver resection, 10 subjects were sacrificed from each group, and liver weights and the mitotic count and Ki-67 were measured. RESULTS Regenerating liver weight on the 1st and 5th days after partial hepatectomy was statistically different in the low dose and high dose nebivolol groups compared to the control group. Mitotic count on the 1st day after partial hepatectomy was significantly higher in the low dose and high dose nebivolol groups than the control group. There was no statistically significant difference detected between the three groups for the 5th day. On the 1st day, Ki-67 rates were significantly higher in both groups given nebivolol than the control group. However, 5th day results were not statistically significant. CONCLUSION Nebivolol increases regeneration after partial hepatectomy in rats.
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Lu C, Xia J, Zhou Y, Lu X, Zhang L, Gou M, Li L, Zhang X, Ji H, Zhu K, Li L, Zhang J, Yu P, Yang J, Bu H, Shi Y. Loss of Gsα impairs liver regeneration through a defect in the crosstalk between cAMP and growth factor signaling. J Hepatol 2016; 64:342-351. [PMID: 26386161 DOI: 10.1016/j.jhep.2015.08.036] [Citation(s) in RCA: 23] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/12/2015] [Revised: 08/17/2015] [Accepted: 08/30/2015] [Indexed: 02/05/2023]
Abstract
BACKGROUND & AIMS The stimulatory G protein α subunit (Gsα) activates the cAMP-dependent pathway by stimulating the production of cAMP and participates in diverse cell processes. Aberrant expression of Gsα results in various pathophysiological disorders, including tumorigenesis, but little is known about its role in liver regeneration. METHODS We generated a hepatocyte-specific Gsα gene knockout mouse to demonstrate the essential role of Gsα in liver regeneration using a mouse model with 70% partial hepatectomy (PH) or an intraperitoneal injection of carbon tetrachloride (CCl4). RESULTS Gsα inactivation dramatically impaired liver regeneration and blocked proliferating hepatocytes in G1/S transition due to the simultaneous depression of cyclin-dependent kinase 2 (CDK2) and cyclin E1. Loss of Gsα led to a fundamental alteration in gene profiles. Among the altered signaling cascades, the MAPK/Erk pathway, which is downstream of growth factor signaling, was disrupted secondary to a defect in phosphorylated Raf1 (pRaf1), resulting in a deficiency in phosphorylated CREB (pCREB) and CDK2 ablation. The lack of pRaf1 also resulted in a failure to phosphorylate retinoblastoma, which releases and activates E2F1, and a decrease in cyclin E1. Although these factors could be phosphorylated through both Gsα and growth factor signaling, the unique function of Raf1 in the growth factor cascade collapsed in response to the lack of Gsα. CONCLUSION The growth factor signaling pathway that promotes hepatocyte proliferation is dependent on Gsα signaling. Loss of Gsα leads to a breakdown of the crosstalk between cAMP and growth factor signaling and dramatically impairs liver regeneration.
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Affiliation(s)
- Changli Lu
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Department of Pathology, West China Hospital, Sichuan University, Chengdu, China
| | - Jie Xia
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China
| | - Yongjie Zhou
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital, Sichuan University, China
| | - Xufeng Lu
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China
| | - Lei Zhang
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital, Sichuan University, China
| | - Maling Gou
- State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, China
| | - Lei Li
- Department of Liver Surgery, West China Hospital, Sichuan University, Chengdu, China
| | - Xiaoyun Zhang
- Department of Liver Surgery, West China Hospital, Sichuan University, Chengdu, China
| | - Hongjie Ji
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital, Sichuan University, China
| | - Keting Zhu
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital, Sichuan University, China
| | - Li Li
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China
| | - Jie Zhang
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China
| | - Ping Yu
- Laboratory of Cell and Gene Therapy, West China Institute of Women and Children's Health, West China Second University Hospital, Sichuan University, Chengdu, China
| | - Jiayin Yang
- Department of Liver Surgery, West China Hospital, Sichuan University, Chengdu, China
| | - Hong Bu
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Department of Pathology, West China Hospital, Sichuan University, Chengdu, China.
| | - Yujun Shi
- Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, China; Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital, Sichuan University, China.
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Jahouh F, Marongiu F, Serra MP, Laconi E, Banoub J. Gas-phase fragmentation of the N-oxide and N-hydroxylated derivatives of retrorsine using liquid chromatography/electrospray ionization quadrupole time-of-flight tandem mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2015; 29:1733-1748. [PMID: 26331923 DOI: 10.1002/rcm.7276] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/13/2015] [Revised: 07/09/2015] [Accepted: 07/11/2015] [Indexed: 06/05/2023]
Abstract
RATIONALE We report the electrospray ionization mass spectrometry and low-energy collision-induced dissociation tandem mass spectrometry (CID-MS/MS) analysis of a pyrrolizidine alkaloid extract containing both retrorsine [C18H25NO6] and its N-oxide [C18H25NO7] and N-hydroxyl [C18H26NO7] derivatives measured with a QqTOFMS hybrid instrument. METHODS A solution of the pyrrolizidine alkaloid extract containing retrorsine and its N-oxide and N-hydroxyl derivatives was directly infused into an electrospray ionization-quadrupole-time-of-flight (ESI-QTOF) mass spectrometer and product ion scans of the protonated molecules of each species were acquired. Labile protons of each compound were deuterated and computational energy calculations of the proposed structures of the product ions were used to determine the fragmentation pathways of retrorsine and its N-oxide and N-hydroxyl derivatives. RESULTS ESI-MS of the pyrrolizidine alkaloid extract containing retrorsine and its N-oxide and N-hydroxyl derivatives afforded the protonated retrorsine [M1 + H](+) at m/z 352.1760 and the protonated retrorsine N-oxide [M2 + H](+) at m/z 368.1631 in addition to the formation of the unexpected protonated N-hydroxyl radical [M3 + H](+•) at m/z 369.1686. CID-MS/MS of this series of protonated molecules allowed the evaluation of their gas-phase fragmentations and the establishment of their fragmentation pathways. It was also found that several product ions could be assigned to different structures. Deuterium exchange and computational energy calculations allowed us to determine the most probable structures for the characterized product ions. CONCLUSIONS To our knowledge, the identification of the protonated retrorsine N-hydroxyl radical [M3 + H](+•) is reported for the first time. In addition, the MS/MS results can be used for the identification of retrorsine and its N-oxide and N-hydroxyl derivatives in different complex biological matrices.
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Affiliation(s)
- Farid Jahouh
- Chemistry Department, Memorial University of Newfoundland, Saint John's, Canada
- Special Projects, Science Branch, Department of Fisheries and Oceans Canada, Saint John's, Canada
| | - Fabio Marongiu
- Department of Biomedical Sciences, University of Cagliari, Italy
| | | | - Ezio Laconi
- Department of Biomedical Sciences, University of Cagliari, Italy
| | - Joseph Banoub
- Chemistry Department, Memorial University of Newfoundland, Saint John's, Canada
- Special Projects, Science Branch, Department of Fisheries and Oceans Canada, Saint John's, Canada
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13
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Sandini TM, Udo MS, Reis‐Silva TM, Sanches D, Bernardi MM, Flório JC, Spinosa HDS. Prenatal exposure to integerrimine N‐oxide enriched butanolic residue from
Senecio brasiliensis
affects behavior and striatal neurotransmitter levels of rats in adulthood. Int J Dev Neurosci 2015; 47:157-64. [DOI: 10.1016/j.ijdevneu.2015.09.003] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2015] [Revised: 09/15/2015] [Accepted: 09/16/2015] [Indexed: 01/07/2023] Open
Affiliation(s)
- Thaísa M. Sandini
- Department of Clinical and Toxicological AnalysesFaculty of Pharmaceutical SciencesUniversity of São PauloAv. Prof. Dr. Lineu Prestes, 58005508‐000São PauloBrazil
| | - Mariana S.B. Udo
- Department of Clinical and Toxicological AnalysesFaculty of Pharmaceutical SciencesUniversity of São PauloAv. Prof. Dr. Lineu Prestes, 58005508‐000São PauloBrazil
| | - Thiago M. Reis‐Silva
- Department of NeuroscienceInstitute of PsychologyUniversity of São PauloAv. Prof. Dr. Melo de Morais, 172105508‐030São PauloBrazil
| | - Daniel Sanches
- Department of PathologySchool of Veterinary MedicineUniversity of Sao PauloAv. Prof. Dr. Orlando Marques de Paiva, 8705508 270São PauloBrazil
| | - Maria Martha Bernardi
- Graduate Program of Environmental and Experimental Pathology and Graduate Program DentistryPaulista University, UNIPRua Dr. Bacelar, 121204026‐002São PauloBrazil
| | - Jorge Camilo Flório
- Department of PathologySchool of Veterinary MedicineUniversity of Sao PauloAv. Prof. Dr. Orlando Marques de Paiva, 8705508 270São PauloBrazil
| | - Helenice de S. Spinosa
- Department of PathologySchool of Veterinary MedicineUniversity of Sao PauloAv. Prof. Dr. Orlando Marques de Paiva, 8705508 270São PauloBrazil
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14
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Marongiu F, Serra MP, Sini M, Angius F, Laconi E. Clearance of senescent hepatocytes in a neoplastic-prone microenvironment delays the emergence of hepatocellular carcinoma. Aging (Albany NY) 2014; 6:26-34. [PMID: 24464501 PMCID: PMC3927807 DOI: 10.18632/aging.100631] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
Increasing evidence indicates that carcinogenesis is dependent on the tissue context in which it occurs, implying that the latter can be a target for preventive or therapeutic strategies. We tested the possibility that re-normalizing a senescent, neoplastic-prone tissue microenvironment would exert a modulatory effect on the emergence of neoplastic disease. Rats were exposed to a protocol for the induction of hepatocellular carcinoma (HCC). Using an orthotopic and syngeneic system for cell transplantation, one group of animal was then delivered 8 million normal hepatocytes, via the portal circulation. Hepatocytes transplantation resulted in a prominent decrease in the incidence of both pre-neoplastic and neoplastic lesions. At the end of 1 year 50% of control animals presented with HCC, while no HCC were observed in the transplanted group. Extensive hepatocyte senescence was induced by the carcinogenic protocol in the host liver; however, senescent cells were largely cleared following infusion of normal hepatocytes. Furthermore, levels of Il-6 increased in rats exposed to the carcinogenic protocol, while they returned to near control values in the group receiving hepatocyte transplantation. These results support the concept that strategies aimed at normalizing a neoplastic-prone tissue landscape can modulate progression of neoplastic disease.
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Affiliation(s)
- Fabio Marongiu
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, 09124 Cagliari, Italy
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15
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Abstract
Despite the tremendous hurdles presented by the complexity of the liver's structure and function, advances in liver physiology, stem cell biology and reprogramming, and the engineering of tissues and devices are accelerating the development of cell-based therapies for treating liver disease and liver failure. This State of the Art Review discusses both the near- and long-term prospects for such cell-based therapies and the unique challenges for clinical translation.
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Affiliation(s)
- Sangeeta N Bhatia
- Institute for Medical Engineering & Science at MIT, Department of Electrical Engineering and Computer Science, David H. Koch Institute at MIT, and the Howard Hughes Medical Institute, Cambridge, MA 02139, USA. Division of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA.
| | - Gregory H Underhill
- Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
| | - Kenneth S Zaret
- Institute for Regenerative Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Ira J Fox
- Department of Surgery, Children's Hospital of Pittsburgh of UPMC, University of Pittsburgh School of Medicine, and McGowan Institute for Regenerative Medicine, Pittsburgh, PA 15224, USA
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16
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Serra MP, Marongiu F, Sini M, Marongiu M, Contini A, Wolff H, Rave-Frank M, Krause P, Laconi E, Koenig S. Hepatocyte senescence induced by radiation and partial hepatectomy in rat liver. Int J Radiat Biol 2014; 90:876-83. [PMID: 24827852 DOI: 10.3109/09553002.2014.922714] [Citation(s) in RCA: 12] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022]
Abstract
PURPOSE Exposure to radiation primes the liver for extensive replacement of the resident parenchymal cells by transplanted hepatocytes. The mechanisms underlying this repopulation remain to be clarified. In these studies, we examined the possible occurrence of cell senescence in vivo following radiation-associated preconditioning of the host liver. MATERIALS AND METHODS Fischer 344 rats underwent external-beam, computed-tomography-based partial liver irradiation. A single dose of 25 Gy was delivered to the right liver lobes (40% of liver mass). An additional group of animals received a 1/3 partial hepatectomy (removal of the left anterior lobe) four days after irradiation. Non-irradiated groups served as controls. All rats were sacrificed four weeks after the initial treatment. RESULTS The irradiated livers displayed several markers of cell senescence, including expression of senescence-associated-β-galactosidase (SA-β-gal), increase in cell size, and up-regulation of cyclin-dependent kinase inhibitors (CDK-I) p16 and p21. Furthermore, quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) analysis revealed activation of the senescence-associated secretory phenotype (SASP), including the cytokines interleukin 6 (IL6) and 1α (IL1α). The senescence-related changes were more prominent in rats undergoing partial hepatectomy (PH) following irradiation (IR). CONCLUSIONS We conclude that priming with radiation for liver repopulation results in the induction of cell senescence and the up-regulation of a senescence-associated secretory phenotype. The latter can contribute to the extensive growth of transplanted cells in this system.
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17
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Abstract
The transplantation of hepatocytes could be an alternative therapeutic option to the whole organ transplantation for the treatment of end-stage liver diseases. However, this cell-based therapy needs the understanding of the molecular mechanisms to improve efficacy. This chapter includes a detailed method of a rat model for liver regeneration studies after age-dependent hepatocyte transplantation.
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Affiliation(s)
- Peggy Stock
- Department of Surgery, University of Leipzig, Liebigstraße 21, 04103, Leipzig, Germany,
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18
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Cilekar M, Uysal O, Bal C, Turel S, Yılmaz S. Leptin increases mitotic index and regeneration ratio in hepatectomized rats. Med Sci Monit Basic Res 2013; 19:279-84. [PMID: 24220642 PMCID: PMC3852623 DOI: 10.12659/msmbr.889591] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022] Open
Abstract
Background The aim of this study was to evaluate the potential effect of intraperitoneal administration of leptin on the hepatic regeneration and the mitotic index. Material/Methods 56 Sprague-Dawley rats were divided into 7 groups each containing 8 rats. Group 1 was evaluated as the sham group and no surgical procedure was performed on animals. The rats in groups 2, 3, and 4 (named C24, C48, C72, respectively) were given intraperitoneal injection of 2 ml/kg normal saline 60 minutes before the surgical procedure consisting of laparotomy and 70% hepatectomy. These groups were used as controls at 24, 48, and 72 hours. The rats in groups 5, 6, and 7 (named L24, L48, and L72, respectively) were given intraperitoneal injection of 20 μg/kg doses of recombinant mouse leptin 60 minutes before the same surgical procedure. These groups were evaluated as the experiment groups at 24, 48, and 72 hours. Blood samples were collected for aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and the remaining tissue samples were obtained for liver histopathology, regeneration rate, and mitotic index (MI). The weights of the remaining livers were also noted. Results The values of AST and ALT were higher in the groups that were administered leptin and they had significantly higher mitotic index than the other groups. Leptin also significantly increased the regeneration ratio as compared to the control group. The weights of the remaining livers were also higher in the leptin groups. Conclusions Intraperitoneal administration of leptin was observed to increase liver regeneration and mitotic rate in 70% hepatectomized rats.
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Affiliation(s)
- Murat Cilekar
- Department of General Surgery, Eskişehir Osmangazi University, Medical Faculty, Eskisehir, Turkey
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19
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Oguz S, Kanter M, Erboga M, Toydemir T, Sayhan MB, Onur H. Effects of Urtica dioica on oxidative stress, proliferation and apoptosis after partial hepatectomy in rats. Toxicol Ind Health 2013; 31:475-84. [DOI: 10.1177/0748233713480211] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
The present study was performed to investigate the effect of Urtica dioica (UD) on liver regeneration after partial hepatectomy (PH) in rats. A total of 24 male Sprague Dawley rats were divided into three groups: sham-operated, PH and PH + UD; each group contains eight animals. The rats in UD-treated groups were given UD oils (2 ml/kg/day) once a day orally for 7 days starting 3 days prior to hepatectomy operation. At day 7 after resection, liver samples were collected. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were estimated in liver homogenates. Moreover, histopathological examination, mitotic index (MI), proliferating cell nuclear antigen labeling, proliferation index (PI), transferase-mediated deoxyuridine triphosphate nick end-labeling assay, apoptotic index (AI) were evaluated at day 7 after hepatectomy. As a result, UD significantly increased MI and PI, significantly decreased AI and also attenuated hepatic vacuolar degeneration and sinusoidal congestion in PH rats. UD treatment significantly decreased the elevated tissue MDA level and increased the reduced SOD activity and GSH level in the tissues. These results suggest that UD pretreatment was beneficial for rat liver regeneration after partial hepatectomy.
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Affiliation(s)
- Serhat Oguz
- Department of General Surgery, Faculty of Medicine, Trakya University, Edirne, Turkey
| | - Mehmet Kanter
- Department of Histology and Embryology, Faculty of Medicine, Istanbul Medeniyet University, Istanbul, Turkey
| | - Mustafa Erboga
- Department of Histology and Embryology, Faculty of Medicine, Namik Kemal University, Tekirdag, Turkey
| | - Toygar Toydemir
- Department of General Surgery, Istanbul Surgical Hospital, Istanbul, Turkey
| | - Mustafa Burak Sayhan
- Department of Emergency Medicine, Faculty of Medicine, Trakya University, Edirne, Turkey
| | - Hatice Onur
- Department of Pediatrics, Istanbul Research and Training Hospital, Istanbul, Turkey
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20
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Pichard V, Couton D, Desdouets C, Ferry N. Polyploidization without mitosis improves in vivo liver transduction with lentiviral vectors. Hum Gene Ther 2013; 24:143-51. [PMID: 23249390 DOI: 10.1089/hum.2011.227] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022] Open
Abstract
Lentiviral vectors are efficient gene delivery vehicles for therapeutic and research applications. In contrast to oncoretroviral vectors, they are able to infect most nonproliferating cells. In the liver, induction of cell proliferation dramatically improved hepatocyte transduction using all types of retroviral vectors. However, the precise relationship between hepatocyte division and transduction efficiency has not been determined yet. Here we compared gene transfer efficiency in the liver after in vivo injection of recombinant lentiviral or Moloney murine leukemia viral (MoMuLV) vectors in hepatectomized rats treated or not with retrorsine, an alkaloid that blocks hepatocyte division and induces megalocytosis. Partial hepatectomy alone resulted in a similar increase in hepatocyte transduction using either vector. In retrorsine-treated and partially hepatectomized rats, transduction with MoMuLV vectors dropped dramatically. In contrast, we observed that retrorsine treatment combined with partial hepatectomy increased lentiviral transduction to higher levels than hepatectomy alone. Analysis of nuclear ploidy in single cells showed that a high level of transduction was associated with polyploidization. In conclusion, endoreplication could be exploited to improve the efficiency of liver-directed lentiviral gene therapy.
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Affiliation(s)
- Virginie Pichard
- INSERM UMR-S 948, Université de Nantes, CHU Hôtel Dieu, 44093 Nantes, France
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21
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Toydemir T, Kanter M, Erboga M, Oguz S, Erenoglu C. Antioxidative, antiapoptotic, and proliferative effect of curcumin on liver regeneration after partial hepatectomy in rats. Toxicol Ind Health 2013; 31:162-72. [PMID: 23299190 DOI: 10.1177/0748233712469658] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
The aim of the present study was to assess the influence of curcumin on liver regeneration after partial hepatectomy (PH) in rats. A total of 24 male Sprague Dawley rats were divided into three groups: sham-operated (SH), PH, and PH + curcumin; each group contains eight animals. The rats in curcumin-treated groups were given curcumin (in a dose of 100 mg/kg body weight) once a day orally for 7 days, starting 3 days prior to hepatectomy operation. At 7 days after resection, liver samples were collected. The malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) levels were estimated in liver homogenates. Moreover, histopathological examination, mitotic index (MI), proliferating cell nuclear antigen labeling, proliferation index (PI), transferase-mediated 2'-deoxyuridine, 5'-triphosphate nick end-labeling assay, and apoptotic index (AI) were evaluated at 7 days after hepatectomy. As a result, curcumin significantly increased MI and PI and significantly decreased AI in PH rats. Additionally, curcumin remarkably inhibited MDA elevation, restored impaired antioxidant SOD activity and GSH level and also attenuated hepatic vacuolar degeneration and sinusoidal congestion. These results suggested that curcumin treatment had a beneficial effect on liver regenerative capacity of the remnant liver tissue after hepatectomy, probably due to its antioxidative, antiapoptotic, and proliferative properties.
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Affiliation(s)
- Toygar Toydemir
- Department of General Surgery, Istanbul Surgery Hospital, Istanbul, Turkey
| | - Mehmet Kanter
- Department of Histology and Embryology, Faculty of Medicine, Istanbul Medeniyet University, Istanbul, Turkey
| | - Mustafa Erboga
- Department of Histology and Embryology, Faculty of Medicine, Namik Kemal University, Tekirdag, Turkey
| | - Serhat Oguz
- Department of General Surgery, Faculty of Medicine, Trakya University, Edirne, Turkey
| | - Cengiz Erenoglu
- Department of General Surgery, Faculty of Medicine, Trakya University, Edirne, Turkey
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22
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Fan L, Xu C, Wang C, Tao J, Ho C, Jiang L, Gui B, Huang S, Evert M, Calvisi DF, Chen X. Bmi1 is required for hepatic progenitor cell expansion and liver tumor development. PLoS One 2012; 7:e46472. [PMID: 23029524 PMCID: PMC3460872 DOI: 10.1371/journal.pone.0046472] [Citation(s) in RCA: 28] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/12/2011] [Accepted: 09/02/2012] [Indexed: 12/13/2022] Open
Abstract
Bmi1 is a polycomb group transcriptional repressor and it has been implicated in regulating self-renewal and proliferation of many types of stem or progenitor cells. In addition, Bmi1 has been shown to function as an oncogene in multiple tumor types. In this study, we investigated the functional significance of Bmi1 in regulating hepatic oval cells, the major type of bipotential progenitor cells in adult liver, as well as the role of Bmi1 during hepatocarcinogenesis using Bmi1 knockout mice. We found that loss of Bmi1 significantly restricted chemically induced oval cell expansion in the mouse liver. Concomitant deletion of Ink4a/Arf in Bmi1 deficient mice completely rescued the oval cell expansion phenotype. Furthermore, ablation of Bmi1 delayed hepatocarcinogenesis induced by AKT and Ras co-expression. This antineoplastic effect was accompanied by the loss of hepatic oval cell marker expression in the liver tumor samples. In summary, our data demonstrated that Bmi1 is required for hepatic oval cell expansion via deregulating the Ink4a/Arf locus in mice. Our study also provides the evidence, for the first time, that Bmi1 expression is required for liver cancer development in vivo, thus representing a promising target for innovative treatments against human liver cancer.
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Affiliation(s)
- Lingling Fan
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
- Center for Stem Cell Research and Application, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Chuanrui Xu
- School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Chunmei Wang
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
| | - Junyan Tao
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
| | - Coral Ho
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
| | - Lijie Jiang
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
| | - Bing Gui
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
| | - Shiang Huang
- Center for Stem Cell Research and Application, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
| | - Matthias Evert
- Institute of Pathology, University of Greifswald, Greifswald, Germany
| | - Diego F. Calvisi
- Institute of Pathology, University of Greifswald, Greifswald, Germany
| | - Xin Chen
- Department of Bioengineering and Therapeutic Sciences, University of California San Francisco, San Francisco, California, United States of America
- Liver Center, University of California San Francisco, San Francisco, California, United States of America
- * E-mail:
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23
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Serra MP, Marongiu F, Sini M, Laconi E. Hepatocyte senescence in vivo following preconditioning for liver repopulation. Hepatology 2012; 56:760-8. [PMID: 22392699 DOI: 10.1002/hep.25698] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/05/2012] [Accepted: 02/23/2012] [Indexed: 12/20/2022]
Abstract
UNLABELLED In the retrorsine (RS)-based model of massive liver repopulation, preexposure to this naturally occurring alkaloid is sufficient to prime normal host parenchymal cells to be slowly replaced by transplanted normal hepatocytes. The basis for this striking effect is yet to be fully elucidated. In the present studies the possible involvement of cell senescence was investigated. Fischer 344 rats were treated according to the RS-based protocol for hepatocyte transplantation, i.e., two doses of RS, 2 weeks apart, and were killed at 4 or 8 weeks after treatment. Control groups were given saline. Expression of senescence-associated beta-galactosidase was greatly induced in hepatocytes exposed to RS. In addition, several other changes that have been related to cell senescence were observed: these included markers of persistent activation of a DNA damage response, an increased expression of mammalian target of rapamycin, and positive regulators of the cell cycle, together with the induction of p21 and p27 cyclin-dependent kinase inhibitors. Furthermore, RS treatment increased levels of interleukin-6 in the liver, consistent with the activation of a senescence-associated secretory phenotype. CONCLUSION These findings indicate that RS induces hepatocyte senescence in vivo. We propose that cell senescence and the associated secretory phenotype can contribute to the selective growth of transplanted hepatocytes in this system.
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Affiliation(s)
- Maria Paola Serra
- Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy
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24
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Christ B, Brückner S. Rodent animal models for surrogate analysis of cell therapy in acute liver failure. Front Physiol 2012; 3:78. [PMID: 22485094 PMCID: PMC3317270 DOI: 10.3389/fphys.2012.00078] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2012] [Accepted: 03/16/2012] [Indexed: 12/27/2022] Open
Abstract
Without therapeutic intervention acute liver failure (ALF) is the consequence of a progredient destruction of the liver parenchyma due to metabolic exhaustion of the hepatocytes. Perivenous hepatocytes are responsible for the detoxification of noxious compounds via the cytochrome P450 enzyme system. Liver transplantation is the only remaining therapeutic option in the end-stage of the disease. Assuming that metabolic capacity could be provided by healthy hepatocytes and thus substitute for the genuine parenchymal cells hepatocyte transplantation since quite some time is considered to be an alternative to whole liver transplantation. While this hypothesis achieved proof-of-concept in animal trials clinical breakthrough is still awaiting success, the reasons of which are ongoing matter of debate. In recent times mesenchymal stem cells (MSC) came into focus as a transplantable cell source to treat ALF. Interestingly, as demonstrated in various rodent animal models their mode of action is rather based on trophic support of hepatocytes remaining in the damaged host parenchyma rather than substitution of tissue loss. Mechanistically, either direct or indirect paracrine effects from the transplanted cells acting pro-proliferative, anti-apoptotic, and anti-inflammatory seem to trigger the regenerative response of the residual healthy hepatocytes in the otherwise lethally injured liver parenchyma. Thus, allogeneic MSC may be the best choice for the treatment of ALF taking advantage of their short-term benefit to sustain the critical phase of the acute insult avoiding long-term immunosuppression.
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Affiliation(s)
- Bruno Christ
- Applied Molecular Hepatology Laboratory, Department of Visceral, Transplantation, Thoracic and Vascular Surgery, University Hospital Leipzig Leipzig, Germany
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25
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Hübner S, Efthymiadis A. Recent progress in histochemistry and cell biology. Histochem Cell Biol 2012; 137:403-57. [PMID: 22366957 DOI: 10.1007/s00418-012-0933-4] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 02/06/2012] [Indexed: 01/06/2023]
Abstract
Studies published in Histochemistry and Cell Biology in the year 2011 represent once more a manifest of established and newly sophisticated techniques being exploited to put tissue- and cell type-specific molecules into a functional context. The review is therefore the Histochemistry and Cell Biology's yearly intention to provide interested readers appropriate summaries of investigations touching the areas of tissue biology, developmental biology, the biology of the immune system, stem cell research, the biology of subcellular compartments, in order to put the message of such studies into natural scientific-/human- and also pathological-relevant correlations.
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Affiliation(s)
- Stefan Hübner
- Institute of Anatomy and Cell Biology, University of Würzburg, Würzburg, Germany.
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26
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Serra MP, Doratiotto S, Marongiu F, Laconi E. Normal hepatocyte transplantation delays the emergence of chemically induced preneoplastic nodules in rat liver. Cell Transplant 2011; 21:671-7. [PMID: 21944459 DOI: 10.3727/096368911x600975] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
Abstract
Cancer often arises in a background of chronic tissue damage. It is also increasingly appreciated that such an injured tissue microenvironment might foster the selective emergence of altered cells, leading to neoplasia. Accordingly, reversal of chronic tissue damage could represent a potential strategy to counteract neoplastic disease. In these studies, we aim to investigate whether transplantation of normal cells in the context of an injured, neoplastic-prone microenvironment might impact on the evolution of the carcinogenic process. A rat model of chemically induced hepatocarcinogenesis was used. Animals were given a single dose of diethylnitrosamine (DENA), followed by two injections of retrorsine (RS), a pyrrolizidine alkaloid that imposes a persistent block on hepatocyte cell cycle. At the end of this protocol, rats were either given no further treatment or injected, via the portal circulation, with 4 million normal hepatocytes isolated from a syngenic donor. After 3 months, rats given DENA+RS alone displayed numerous discrete nodular lesions (up to 30 per liver), ranging 1 to 3 mm in size. On the other hand, in animals receiving DENA+RS and transplantation, donor hepatocytes were able to repopulate over 50% of the host liver, as expected. Most importantly, both the number and the size of hepatocyte nodules were greatly reduced in these animals (percent nodular area was 1.8 ± 0.3, down from a control value of 8.5 ± 2.8). The above data indicate that strategies aimed at reestablishing a normal tissue microenvironment might be relevant to the management of neoplastic disease.
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Affiliation(s)
- Maria Paola Serra
- Department of Biomedical Sciences, Unit of Experimental Medicine, University of Cagliari, Cagliari, Italy
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27
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Shi Y, Sun H, Bao J, Zhou P, Zhang J, Li L, Bu H. Activation of inactive hepatocytes through histone acetylation: a mechanism for functional compensation after massive loss of hepatocytes. THE AMERICAN JOURNAL OF PATHOLOGY 2011; 179:1138-1147. [PMID: 21763259 PMCID: PMC3157283 DOI: 10.1016/j.ajpath.2011.05.029] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/14/2010] [Revised: 05/11/2011] [Accepted: 05/16/2011] [Indexed: 02/05/2023]
Abstract
The mechanisms by which hepatic function is maintained after extensive parenchymal loss are unclear. In this study, we propose a novel concept of "functional heterogeneity" of hepatocytes based on their different expression of acetylated histones, the markers of active gene transcription, to explain the powerful compensatory capability of the liver. In the healthy human liver, only a fraction of the hepatocytes were marked by acetylated histones (ac-H2AK5, ac-H2BK5, ac-H3K9, ac-H3K14, ac-H3K27, and ac-H3K9/14). With the progression of cirrhosis, the ratio of the positive cells was gradually elevated, accompanied by the gradual exhaustion of the negative cells. By examining the global transcriptome of the mouse hepatocytes, we observed that the primed genes in the positive cells were much more numerous than those in negative cells. In a 70% hepatectomized mouse, the remnant hepatocytes were extensively activated, and the liver function was well maintained even when regeneration was severely inhibited. The functional compensation was absolutely dependent on the elevated expression of acetyl-histones. Additionally, when liver regeneration was blocked, the metabolism-related genes seemed to be preferentially transcribed. In conclusion, we demonstrate that normally, part of the active hepatocytes are competent for routine physiological requirements. The inactive hepatocytes, delicately regulated by acetyl-histones, act as a functional reservoir for future activation to restore the liver function after massive parenchymal loss.
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Affiliation(s)
- Yujun Shi
- Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, China
| | - Huaiqiang Sun
- Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, China
| | - Ji Bao
- Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, China
| | - Ping Zhou
- Department of Pathology, West China Hospital, Sichuan University, Chengdu, China
| | - Jie Zhang
- Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, China
| | - Li Li
- Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, China
| | - Hong Bu
- Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, China
- Department of Pathology, West China Hospital, Sichuan University, Chengdu, China
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28
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Doratiotto S, Krause P, Serra MP, Marongiu F, Sini M, Koenig S, Laconi E. The growth pattern of transplanted normal and nodular hepatocytes. Histochem Cell Biol 2011; 135:581-91. [PMID: 21528371 PMCID: PMC3106155 DOI: 10.1007/s00418-011-0813-3] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 04/15/2011] [Indexed: 01/16/2023]
Abstract
Overt neoplasia is often the end result of a long biological process beginning with the appearance of focal lesions of altered tissue morphology. While the putative clonal nature of focal lesions has often been emphasized, increasing attention is being devoted to the possible role of an altered growth pattern in the evolution of carcinogenesis. Here we compare the growth patterns of normal and nodular hepatocytes in a transplantation system that allows their selective clonal proliferation in vivo. Rats were pre-treated with retrorsine, which blocks the growth of resident hepatocytes, and were then transplanted with hepatocytes isolated from either normal liver or hepatocyte nodules. Both cell types were able to proliferate extensively in the recipient liver, as expected. However, their growth pattern was remarkably different. Clusters of normal hepatocytes integrated in the host liver, displaying a normal histology; however, transplanted nodular hepatocytes formed new hepatocyte nodules, with altered morphology and sharp demarcation from surrounding host liver. Both the expression and distribution of proteins involved in cell polarity, cell communication, and cell adhesion, including connexin 32, E-cadherin, and matrix metalloproteinase-2, were altered in clusters of nodular hepatocytes. Furthermore, we were able to show that down-regulation of connexin 32 and E-cadherin in nodular hepatocyte clusters was independent of growth rate. These results support the concept that a dominant pathway towards neoplastic disease in several organs involves defect(s) in tissue pattern formation.
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Affiliation(s)
- Silvia Doratiotto
- Department of Biomedical Sciences and Biotechnology, Section of Experimental Pathology, University of Cagliari, Via Porcell, 4, 09125 Cagliari, Italy
| | - Petra Krause
- Department of General Surgery, University Medical Centre Goettingen, Göttingen, Germany
| | - Maria Paola Serra
- Department of Biomedical Sciences and Biotechnology, Section of Experimental Pathology, University of Cagliari, Via Porcell, 4, 09125 Cagliari, Italy
| | - Fabio Marongiu
- Department of Biomedical Sciences and Biotechnology, Section of Experimental Pathology, University of Cagliari, Via Porcell, 4, 09125 Cagliari, Italy
| | - Marcella Sini
- Department of Biomedical Sciences and Biotechnology, Section of Experimental Pathology, University of Cagliari, Via Porcell, 4, 09125 Cagliari, Italy
| | - Sarah Koenig
- Department of General Surgery, University Medical Centre Goettingen, Göttingen, Germany
| | - Ezio Laconi
- Department of Biomedical Sciences and Biotechnology, Section of Experimental Pathology, University of Cagliari, Via Porcell, 4, 09125 Cagliari, Italy
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Marongiu F, Gramignoli R, Dorko K, Miki T, Ranade AR, Serra MP, Doratiotto S, Sini M, Sharma S, Mitamura K, Sellaro TL, Tahan V, Skvorak KJ, Ellis EC, Badylak SF, Davila JC, Hines R, Laconi E, Strom SC. Hepatic differentiation of amniotic epithelial cells. Hepatology 2011; 53:1719-29. [PMID: 21374689 PMCID: PMC3103747 DOI: 10.1002/hep.24255] [Citation(s) in RCA: 104] [Impact Index Per Article: 7.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
UNLABELLED Hepatocyte transplantation to treat liver disease is largely limited by the availability of useful cells. Human amniotic epithelial cells (hAECs) from term placenta express surface markers and gene characteristics of embryonic stem cells and have the ability to differentiate into all three germ layers, including tissues of endodermal origin (i.e., liver). Thus, hAECs could provide a source of stem cell-derived hepatocytes for transplantation. We investigated the differentiation of hAECs in vitro and after transplantation into the livers of severe combined immunodeficient (SCID)/beige mice. Moreover, we tested the ability of rat amniotic epithelial cells (rAECs) to replicate and differentiate upon transplantation into a syngenic model of liver repopulation. In vitro results indicate that the presence of extracellular matrix proteins together with a mixture of growth factors, cytokines, and hormones are required for differentiation of hAECs into hepatocyte-like cells. Differentiated hAECs expressed hepatocyte markers at levels comparable to those of fetal hepatocytes. They were able to metabolize ammonia, testosterone, and 17α-hydroxyprogesterone caproate, and expressed inducible fetal cytochromes. After transplantation into the liver of retrorsine (RS)-treated SCID/beige mice, naïve hAECs differentiated into hepatocyte-like cells that expressed mature liver genes such as cytochromes, plasma proteins, transporters, and other hepatic enzymes at levels equal to adult liver tissue. When transplanted in a syngenic animal pretreated with RS, rAECs were able to engraft and generate a progeny of cells with morphology and protein expression typical of mature hepatocytes. CONCLUSION Amniotic epithelial cells possess the ability to differentiate into cells with characteristics of functional hepatocytes both in vitro and in vivo, thus representing a useful and noncontroversial source of cells for transplantation.
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Affiliation(s)
- Fabio Marongiu
- Department of Pathology, University of Pittsburgh, PA, USA
- Department of Biomedical Sciences and Technologies, Università degli Studi di Cagliari, Italy
| | | | - Kenneth Dorko
- Department of Pathology, University of Pittsburgh, PA, USA
| | - Toshio Miki
- Department of Pathology, University of Pittsburgh, PA, USA
- McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA, USA
| | | | - Maria Paola Serra
- Department of Biomedical Sciences and Technologies, Università degli Studi di Cagliari, Italy
| | - Silvia Doratiotto
- Department of Biomedical Sciences and Technologies, Università degli Studi di Cagliari, Italy
| | - Marcella Sini
- Department of Biomedical Sciences and Technologies, Università degli Studi di Cagliari, Italy
| | - Shringi Sharma
- Department of Pharmaceutical Sciences, University of Pittsburgh, PA, USA
| | | | - Tiffany L. Sellaro
- McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA, USA
| | - Veysel Tahan
- Department of Pathology, University of Pittsburgh, PA, USA
| | | | - Ewa C.S. Ellis
- Department of Pathology, University of Pittsburgh, PA, USA
- Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
| | - Stephen F. Badylak
- McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA, USA
| | - Julio C. Davila
- Pfizer Inc., Pfizer Global Research and Development, St. Louis, MO, USA
| | - Ronald Hines
- Children’s Research Institute, Medical College of Wisconsin, Milwaukee, WI, USA
| | - Ezio Laconi
- Department of Biomedical Sciences and Technologies, Università degli Studi di Cagliari, Italy
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Sun YL, Yin SY, Zhou L, Xie HY, Zhang F, Wu LM, Zheng SS. Hepatocyte differentiation of human fibroblasts from cirrhotic liver in vitro and in vivo. Hepatobiliary Pancreat Dis Int 2011; 10:55-63. [PMID: 21269936 DOI: 10.1016/s1499-3872(11)60008-8] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
BACKGROUND Mesenchymal stem cells (MSCs) and fibroblasts have intimate relationships, and the phenotypic homology between fibroblasts and MSCs has been recently described. The aim of this study was to investigate the hepatic differentiating potential of human fibroblasts in cirrhotic liver. METHODS The phenotypes of fibroblasts in cirrhotic liver were labeled by biological methods. After that, the differentiation potential of these fibroblasts in vitro was characterized in terms of liver-specific gene and protein expression. Finally, an animal model of hepatocyte regeneration in severe combined immunodeficient (SCID) mice was created by retrorsine injection and partial hepatectomy, and the expression of human hepatocyte proteins in SCID mouse livers was checked by immunohistochemical analysis after fibroblast administration. RESULTS Surface immunophenotyping revealed that a minority of fibroblasts expressed markers of MSCs and hepatic epithelial cytokeratins as well as alpha-smooth muscle actin, but homogeneously expressed vimentin, desmin, prolyl 4-hydroxylase and fibronectin. These fibroblasts presented the characteristics of hepatocytes in vitro and differentiated directly into functional hepatocytes in the liver of hepatectomized SCID mice. CONCLUSIONS This study demonstrated that fibroblasts in cirrhotic liver have the potential to differentiate into hepatocyte-like cells in vitro and in vivo. Our findings infer that hepatic differentiation of fibroblasts may serve as a new target for reversion of liver fibrosis and a cell source for tissue engineering.
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Affiliation(s)
- Yu-Ling Sun
- Department of Hepatobiliary and Pancreatic Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China
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Abstract
Mammalian liver has a unique capacity to regenerate following resection or injury, and recovery of liver mass is mainly through proliferation of remaining adult hepatocytes. However, in pathologic conditions, especially during acute liver failure (ALF) and advanced stages of chronic liver disease (CLD), regeneration eventually fails and orthothopic liver transplantation (OLT) represents the only curative approach. The clinical scenario of a world-wide increasing incidence of end-stage CLD and an associated lack of organ availability has led several laboratories to explore the feasibility and efficiency of experimental alternatives to OLT involving cellular therapy. This review presents experimental and clinical studies performed in the last 10-15 years where adult and embryonic hepatocytes, hepatic stem/progenitor cells and extrahepatic stem cells have been used as transplantable cell sources.
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Affiliation(s)
- Francesco Paolo Russo
- Department of Surgical and Gastroenterological Sciences, Gastroenterology Unit, University of Padova, Padova, Italy
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Shupe T, Petersen BE. Potential applications for cell regulatory factors in liver progenitor cell therapy. Int J Biochem Cell Biol 2011; 43:214-21. [PMID: 20851776 PMCID: PMC3022095 DOI: 10.1016/j.biocel.2010.09.005] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/28/2009] [Revised: 08/24/2010] [Accepted: 09/06/2010] [Indexed: 12/18/2022]
Abstract
Orthotopic liver transplant represent the state of the art treatment for terminal liver pathologies such as cirrhosis in adults and hemochromatosis in neonates. A limited supply of transplantable organs in relationship to the demand means that many patients will succumb to disease before an organ becomes available. One promising alternative to liver transplant is therapy based on the transplant of liver progenitor cells. These cells may be derived from the patient, expanded in vitro, and transplanted back to the diseased liver. Inborn metabolic disorders represent the most attractive target for liver progenitor cell therapy, as many of these disorders may be corrected by repopulation of only a portion of the liver by healthy cells. Another potential application for liver progenitor cell therapy is the seeding of bio-artificial liver matrix. These ex vivo bioreactors may someday be used to bridge critically ill patients to other treatments. Conferring a selective growth advantage to the progenitor cell population remains an obstacle to therapy development. Understanding the molecular signaling mechanisms and micro-environmental cues that govern liver progenitor cell phenotype may someday lead to strategies for providing this selective growth advantage. The discovery of a population of cells within the bone marrow possessing the ability to differentiate into hepatocytes may provide an easily accessible source of cells for liver therapies.
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Affiliation(s)
- Thomas Shupe
- Department of Pathology, Immunology and Laboratory Medicine, University of Florida, College of Medicine, Gainesville, FL 32610-0275, USA.
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Christ B, Brückner S, Stock P. Hepatic transplantation of mesenchymal stem cells in rodent animal models. Methods Mol Biol 2011; 698:315-30. [PMID: 21431529 DOI: 10.1007/978-1-60761-999-4_24] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022]
Abstract
The hepatocyte is the smallest functional entity of the liver and executes the majority of this organ's -metabolic functions. Hence, hepatocyte transplantation has become a versatile alternative to whole organ liver transplantation. This novel treatment option is based on the assumption that transplanted -hepatocytes integrate into the host liver, proliferate at the site of tissue damage, take over the long-term hepatic -synthetic capacity, and thus substitute for the diseased host tissue. However, clinical success is still waiting for a breakthrough, likely because of two major reasons including (1) the scarcity of cadaveric donor livers and (2) the largely poor quality of cells isolated from marginal quality donor organs. Therefore, alternative cell sources have to be established to further prompt the clinical success of hepatocyte transplantation. Due to their multiple differentiation potential and nearly unlimited availability, stem cells are an attractive -alternate resource. Because of both clinical and ethical objections, adult stem cells are often preferred over embryonic stem cells as a starting material. Recent studies have demonstrated the ability of mesenchymal stem cells derived from various tissues to differentiate into hepatocyte-like cells in vitro as well as showing specific hepatocyte functions in vivo after transplantation into the livers of mice or rats.
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Affiliation(s)
- Bruno Christ
- First Department of Medicine, Martin-Luther University of Halle-Wittenberg, Halle/Saale, Germany.
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Best DH, Coleman WB. Liver regeneration by small hepatocyte-like progenitor cells after necrotic injury by carbon tetrachloride in retrorsine-exposed rats. Exp Mol Pathol 2010; 89:92-8. [PMID: 20599936 DOI: 10.1016/j.yexmp.2010.06.007] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2010] [Accepted: 06/25/2010] [Indexed: 11/19/2022]
Abstract
Liver regeneration after partial hepatectomy (PH) in rats exposed to the pyrrolizidine alkaloid retrorsine is accomplished through the proliferation and differentiation of a population of small hepatocyte-like progenitor cells (SHPCs). The activation, emergence, and outgrowth of SHPCs in response to the liver deficit generated through surgical PH have been well characterized. However, the participation of these cells in the restoration of hepatocyte numbers and regeneration of liver tissue mass following necrotic injury has not been investigated. To investigate the capacity of SHPCs to respond to necrotizing liver injury, we combined retrorsine treatment with the centrilobular-specific toxin carbon tetrachloride (CCl(4)). Male Fischer 344 rats were treated with retrorsine (30 mg/kg ip) at 6 and 8 weeks of age, followed by CCl(4) treatment (1500 mg/kg ip) 5 weeks later. Liver tissues were harvested at 3, 7, 14, 21, and 30-days post-injection. The dose of CCl(4) employed resulted in the necrotic destruction of 59±2% of liver mass and elicited a regenerative response equivalent to that of surgical PH. Livers from retrorsine-exposed CCl(4)-treated rats exhibit SHPC proliferation similar to retrorsine-exposed rats subjected to PH (RP). SHPCs appear at 3-days post-injection, continue to expand at 7-days and 14-days post-injection, and completely regenerate/restore the liver mass and structure in these animals by 30-days post-injection. The magnitude of SHPC response observed in the undamaged periportal zone of the liver in these animals is unaffected (versus RP rats) by the loss of the centrilobular region. The results of this study show that SHPCs are capable of regenerating liver after exposure to necrotizing agents and suggest that the progenitor cell of origin of the SHPCs is not restricted to the centrilobular zone of the liver parenchyma.
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Affiliation(s)
- D Hunter Best
- Department of Pathology and Laboratory Medicine, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA.
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Gomes MFPL, de Oliveira Massoco C, Xavier JG, Bonamin LV. Comfrey (Symphytum Officinale. l.) and Experimental Hepatic Carcinogenesis: A Short-term Carcinogenesis Model Study. EVIDENCE-BASED COMPLEMENTARY AND ALTERNATIVE MEDICINE : ECAM 2010; 7:197-202. [PMID: 18955295 PMCID: PMC2862927 DOI: 10.1093/ecam/nem172] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 06/10/2005] [Accepted: 09/25/2007] [Indexed: 02/05/2023]
Abstract
Comfrey or Symphytum officinale (L.) (Boraginaceae) is a very popular plant used for therapeutic purposes. Since the 1980s, its effects have been studied in long-term carcinogenesis studies, in which Comfrey extract is administered at high doses during several months and the neoplastic hepatic lesions are evaluated. However, the literature on this topic is very poor considering the studies performed under short-term carcinogenesis protocols, such as the 'resistant hepatocyte model' (RHM). In these studies, it is possible to observe easily the phenomena related to the early phases of tumor development, since pre-neoplastic lesions (PNLs) rise in about 1-2 months of chemical induction. Herein, the effects of chronic oral treatment of rats with 10% Comfrey ethanolic extract were evaluated in a RHM. Wistar rats were sequentially treated with N-nitrosodiethylamine (ip) and 2-acetilaminofluorene (po), and submitted to hepatectomy to induce carcinogenesis promotion. Macroscopic/microscopic quantitative analysis of PNL was performed. Non-parametric statistical tests (Mann-Whitney and χ(2)) were used, and the level of significance was set at P ≤ 0.05. Comfrey treatment reduced the number of pre-neoplastic macroscopic lesions up to 1 mm (P ≤ 0.05), the percentage of oval cells (P = 0.0001) and mitotic figures (P = 0.007), as well as the number of Proliferating Cell Nuclear Antigen (PCNA) positive cells (P = 0.0001) and acidophilic pre-neoplastic nodules (P = 0.05). On the other hand, the percentage of cells presenting megalocytosis (P = 0.0001) and vacuolar degeneration (P = 0.0001) was increased. Scores of fibrosis, glycogen stores and the number of nucleolus organizing regions were not altered. The study indicated that oral treatment of rats with 10% Comfrey alcoholic extract reduced cell proliferation in this model.
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Affiliation(s)
- Maria Fernanda Pereira Lavieri Gomes
- Laboratory of Veterinary Pathology, Faculty of Veterinary Medicine, University of Santo Amaro, Oncocell biotecnologia LTDA and Laboratory of Pathology, Health Sciences Institute, University Paulista, São Paulo, Brazil
| | - Cristina de Oliveira Massoco
- Laboratory of Veterinary Pathology, Faculty of Veterinary Medicine, University of Santo Amaro, Oncocell biotecnologia LTDA and Laboratory of Pathology, Health Sciences Institute, University Paulista, São Paulo, Brazil
| | - José Guilherme Xavier
- Laboratory of Veterinary Pathology, Faculty of Veterinary Medicine, University of Santo Amaro, Oncocell biotecnologia LTDA and Laboratory of Pathology, Health Sciences Institute, University Paulista, São Paulo, Brazil
| | - Leoni Villano Bonamin
- Laboratory of Veterinary Pathology, Faculty of Veterinary Medicine, University of Santo Amaro, Oncocell biotecnologia LTDA and Laboratory of Pathology, Health Sciences Institute, University Paulista, São Paulo, Brazil
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Tschaharganeh DF, Kaldenbach M, Erschfeld S, Tischendorf JJW, Trautwein C, Streetz KL. Glycoprotein 130-dependent pathways in host hepatocytes are important for liver repopulation in mice. Liver Transpl 2010; 16:23-32. [PMID: 20035522 DOI: 10.1002/lt.21962] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
Hepatocyte transplantation (HT) is still restricted by the limited amount of transplantable cells. Therefore, a better understanding of the mechanisms involved in cellular engraftment, proliferation, and in vivo selection is important. Here we aimed to evaluate the role of the interleukin 6 (IL-6)/glycoprotein 130 (gp130) system for liver repopulation. Mice carrying a conditional hepatocyte-specific deletion of the common IL-6 signal transducer gp130 (gp130(Deltahepa)) were used for HT. First, we compared bone marrow transplantation (BMT), partial hepatectomy (PH), and retrorsine treatment of recipient mice to optimize the in vivo selection of transplanted hepatocytes. BMT combined with PH was sufficient to induce a 30-fold increase in the number of transplanted donor hepatocytes, whereas additional retrorsine pretreatment led to an up to 40-fold increase. Next, the influence of gp130 signaling in hepatocytes on cell selection was evaluated. Wild-type (WT) hepatocytes repopulated WT recipients at the same rate as gp130(Deltahepa) cells. In contrast, liver repopulation by transplanted cells was enhanced in gp130(Deltahepa) recipient mice. This was associated with higher proliferation of donor hepatocytes and enhanced apoptosis in gp130(Deltahepa) recipient livers. Additionally, the acute phase response was strongly induced after HT in WT recipients but blunted in gp130(Deltahepa) recipients. As a result, significantly more liver remodeling, evidenced by stronger hepatic stellate cell activation and collagen accumulation, was found in gp130(Deltahepa) mice after HT. In conclusion, the HT model established here can be efficiently applied to investigate cell-specific mechanisms in liver repopulation. Moreover, we have shown that gp130-dependent pathways in host hepatocytes are important for controlling liver repopulation.
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Best DH, Butz GM, Coleman WB. Cytokine-dependent activation of small hepatocyte-like progenitor cells in retrorsine-induced rat liver injury. Exp Mol Pathol 2009; 88:7-14. [PMID: 19874816 DOI: 10.1016/j.yexmp.2009.10.009] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/20/2009] [Accepted: 10/20/2009] [Indexed: 12/22/2022]
Abstract
Complete liver regeneration after partial hepatectomy (PH) in rats exposed to the pyrrolizidine alkaloid retrorsine is accomplished through the activation, expansion, and differentiation of a population of small hepatocyte-like progenitor cells (SHPCs). The mechanism(s) governing activation of SHPCs after PH in retrorsine-injured rats has not been investigated. We examined the possibility that SHPCs require cytokine priming prior to becoming growth factor responsive in this model of liver injury and regeneration. Male Fischer 344 rats were treated with retrorsine (30 mg/kg ip) at 6 and 8 weeks of age. Retrorsine-exposed and age-matched control rats were randomized into dexamethasone-treated and no DEX groups. DEX-treated animals were either given a single dose of DEX (2 mg/kg ip) at the time of PH or multiple DEX treatments (2 mg/kg ip each) at 24 and 1 h before PH and 1, 2, and 3 days post-PH. A subset of rats received 10 microg of recombinant IL6 protein, administered intravenously 30 min after PH. Liver tissues were harvested at 7, 14, 21, and 30 days post-PH. Treatment of retrorsine-exposed rats with the cytokine inhibitor dexamethasone (DEX) effectively blocked the emergence of SHPCs resulting in an inhibition of liver regeneration and producing significant short-term mortality. The livers of DEX-treated retrorsine-exposed rats displayed decreased numbers and smaller SHPC clusters compared to retrorsine-exposed rats in the absence of DEX treatment. Administration of recombinant IL6 to DEX-treated retrorsine-exposed rats restored the emergence of SHPCs and SHPC-mediated regenerative response. The livers of DEX-treated retrorsine-exposed rats that received IL6 displayed numbers of expanding SHPC clusters comparable to that of retrorsine-exposed rats in the absence of DEX treatment. These results combine to suggest that SHPC activation after PH in retrorsine-exposed rats is cytokine dependent and may specifically require IL6.
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Affiliation(s)
- D Hunter Best
- Department of Pathology and Laboratory Medicine, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA.
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Tateno C, Carreiro MP, Hixson DC. Endogenous and transplanted small hepatocytes in retrorsine-treated/partially hepatectomized rat liver show differences in growth, phenotype, and proximity to clusters of gamma-glutamyl transpeptidase-positive host hepatocytes. J Histochem Cytochem 2009; 58:61-72. [PMID: 19786612 DOI: 10.1369/jhc.2009.954560] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
In the present report, we have compared the phenotype and growth of small hepatocyte progenitors (SHPs) induced by retrorsine/partial hepatectomy (R/PH) and small hepatocytes (SHs) isolated from normal adult liver. SHs were isolated by a combination of differential centrifugation and Percoll isodensity fractionation from a liver cell suspension prepared by collagenase perfusion of a dipeptidyl peptidase IV (DPPIV)-positive Fischer F344 rat liver. Following further purification by flow cytometry, the SH-R3 fraction was transplanted via the portal vein into R/PH-treated, DPPIV-negative Fischer F344 rats. Frozen sections from tissue harvested at 5, 7, and 21 days after transplantation were analyzed by indirect immunofluorescence to compare the phenotypic characteristics of colonies formed by exogenous SH-R3s and endogenous SHPs. Colonies of transplanted SHs and endogenous SHPs displayed similar histologies and phenotypes but were distinguished from surrounding hepatocytes by their elevated expression of transferrin receptor. SH-R3 colonies were frequently located within clusters of gamma-glutamyl transpeptidase-positive host hepatocytes. Although significantly smaller at 5 and 7 days after PH, by day 21, SH-R3 colonies were similar in size to those formed by SHPs. The present results suggest that endogenous SHPs are derived, at least in part, from SHPs.
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Affiliation(s)
- Chise Tateno
- Department of Medicine, Division of Hematology and Oncology, Rhode Island Hospital and Brown University Medical School, Providence, Rhode Island 02903, USA
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Brilliant KE, Mills DR, Callanan HM, Hixson DC. Engraftment of syngeneic and allogeneic endothelial cells, hepatocytes and cholangiocytes into partially hepatectomized rats previously treated with mitomycin C. Transplantation 2009; 88:486-95. [PMID: 19696631 PMCID: PMC2730590 DOI: 10.1097/tp.0b013e3181b0b98a] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/27/2023]
Abstract
BACKGROUND Pretreatment with retrorsine crosslinks host hepatocyte DNA and prevents proliferation after partial hepatectomy (PH), allowing selective expansion of transplanted progenitors. Shortcomings are length of protocol and carcinogenicity of retrorsine. METHODS This report describes a rapid liver repopulation protocol using mitomycin C (MMC) to block proliferation of rat hepatocytes in response to PH. One week post-MMC treatment, dipeptidyl peptidase IV negative host rats were given a PH followed by injection of late gestation, newborn, or adult total liver isolates from dipeptidyl peptidase IV positive rats. For allogeneic transplantation, host rats received injections of anti-CD3 antibody before and after PH. RESULTS Host liver staining 2 to 9 weeks posttransplantation revealed well-defined donor hepatocyte colonies with strong canalicular dipeptidyl peptidase IV activity. At the same cell dose, fetal and newborn isolates produced more colonies than adult liver isolates. Hepatocyte colonies also coexpressed marker proteins characteristic of adult hepatocytes and showed polarized localization of plasma membrane proteins. Host livers contained large clusters of sinusoids lined by dipeptidyl peptidase IV positive endothelial cells coexpressing the endothelial cell marker, RECA-1, but lacked the canalicular marker leucine aminopeptidase. Colonies containing donor hepatocytes, endothelial cells, and bile ducts were also observed. Similar levels of engraftment and expansion were achieved with allogeneic liver cell isolates by using anti-CD3 antibody treatment. CONCLUSIONS The MMC transplantation model provides a rapid method for engraftment and expansion of hepatocytes, endothelial cells, and cholangiocytes and should be applicable to investigations centering on the role of endothelial cells in liver regeneration and the identification and characterization of putative endothelial, hepatocyte, and cholangiocyte progenitors.
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Affiliation(s)
- Kate E Brilliant
- Department of Medicine, Division of Hematology and Oncology, Rhode Island Hospital, Providence, RI 02903, USA
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40
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Koenig S, Krause P, Schmidt TK, Rave-Fraenk M, Rothe H, Hermann RM, Becker H, Hess CF, Christiansen H. Irradiation as preparative regimen for hepatocyte transplantation causes prolonged cell cycle block. Int J Radiat Biol 2009; 84:285-98. [DOI: 10.1080/09553000801953359] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/28/2022]
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Zhang B, Inagaki M, Jiang B, Miyakoshi M, Arikura J, Ogawa K, Kasai S. Effects of bone marrow and hepatocyte transplantation on liver injury. J Surg Res 2009; 157:71-80. [PMID: 19345373 DOI: 10.1016/j.jss.2008.12.013] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2008] [Revised: 11/18/2008] [Accepted: 09/01/2008] [Indexed: 01/11/2023]
Abstract
BACKGROUND The therapeutic effects of bone marrow and hepatocyte transplantation were investigated regarding the treatment of retrorsine-partial hepatectomy-induced liver injury. METHODS Analbuminemic F344alb rats were given two doses of retrorsine 2 wk apart, followed 4 wk later by transplantation with F344 rat bone marrow cells or hepatocytes immediately after a two-thirds hepatectomy. The survival rate, liver regeneration rate, liver functions, albumin-positive hepatocytes, and normal albumin gene sequences in the liver and serum albumin levels were investigated in the recipients. RESULTS Although 65% retrorsine/partial hepatectomy-treated F344alb died between 1 and 11 d after the partial hepatectomy, only 27.5% of the animals died following bone marrow transplantation, and 50% with hepatocyte transplantation. Both bone marrow and hepatocyte transplantation ameliorated acute liver injury after a partial hepatectomy. Bone marrow transplantation yielded a very small increase in the number of albumin-positive hepatocytes in the liver, while hepatocyte transplantation resulted in massive replacement of the liver tissues by the donor hepatocytes associated with an elevation of serum albumin after an extended time. CONCLUSIONS Both bone marrow and hepatocyte transplantation could prevent acute hepatic injury, conceivably due to a paracrine mechanism.
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Affiliation(s)
- Biao Zhang
- Department of Surgery, Asahikawa Medical College, Japan
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Ellor S, Shupe T, Petersen B. Stem cell therapy for inherited metabolic disorders of the liver. Exp Hematol 2008; 36:716-725. [PMID: 18375039 PMCID: PMC2443696 DOI: 10.1016/j.exphem.2008.02.002] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/20/2007] [Revised: 01/28/2008] [Accepted: 02/06/2008] [Indexed: 12/16/2022]
Abstract
Modern medicine has conquered an enormous spectrum of health concerns, from the neonatal to the geriatric, the chronically ill to the acutely injured. Among the unmet challenges remaining in modern medicine are inborn disorders of metabolism within the liver. Such inherited metabolic disorders (IMDs) often leave an otherwise healthy individual with a crippling imbalance. As the principal regulator of the body's many metabolic pathways, malencoded hepatic enzymes can drastically disrupt homeostasis throughout the entire body. Severe phenotypes are usually detected within the first few days of life, and treatments range from palliative lifestyle modifications to aggressive surgical procedures. While orthotopic liver transplantation is the single last resort "cure" for these conditions, research during the past few years has brought new therapeutic technologies ever closer to the clinic. Stem cells, therapeutic viral vectors, or a combination thereof, are projected to be the next, best, and final cure for IMDs, which is well-reflected by this generation's research initiatives.
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Affiliation(s)
- Susan Ellor
- Department of Pathology, Immunology and Laboratory Medicine; University of Florida
- The Program for Stem Cell Biology and Regenerative Medicine; University of Florida
| | - Thomas Shupe
- Department of Pathology, Immunology and Laboratory Medicine; University of Florida
- The Program for Stem Cell Biology and Regenerative Medicine; University of Florida
| | - Bryon Petersen
- Department of Pathology, Immunology and Laboratory Medicine; University of Florida
- The Program for Stem Cell Biology and Regenerative Medicine; University of Florida
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Lin H, Mao Q, Wang YM, Jiang L. Proliferation of L02 human hepatocytes in tolerized genetically immunocompetent rats. World J Gastroenterol 2008; 14:2329-37. [PMID: 18416458 PMCID: PMC2705086 DOI: 10.3748/wjg.14.2329] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To investigate whether human hepatocytes could proliferate after transplantation to normal immunocompetent rats treated with 2-acetaminofluorene or Retrorsine and partial hepatectomy.
METHODS: L02 hepatocyte-tolerant Sprague-Dawley rats were injected with Retrorsine, 2-acetaminofluorene or normal saline. L02 hepatocytes were then transplanted via the spleen. Human albumin and its mRNA, specific proliferating cell nuclear antigen (PCNA), L02 hepatocyte dynamic distribution, number density and area density of PCNA-positive cells in the liver were determined.
RESULTS: All the examined indicators were not significantly different between the rats treated with 2-acetaminofluorene and normal saline, which was not the case with rats treated with Retrorsine. A dynamic distribution of L02 hepatocytes in the rat liver was detected from wk 1 to mo 6 after transplantation in the Retrorsine group and from wk 1 to 10 in the 2-acetaminofluorene group. Human albumin and its mRNA were detected from wk 2 to mo 6 in the Retrorsine group and from wk 1 to 8 in the 2-acetaminofluorene group. Specific human PCNA was detected in the rat liver from wk 2 to mo 6 in the Retrorsine group and from wk 2 to 6 in the 2-acetaminofluorene group. Human albumin and its mRNA contents as well as the number of PCNA positive cells reached a peak at wk 4.
CONCLUSION: L02 human hepatocytes could not proliferate significiantly after transplantation to the normal, immunocompetent rats treated with 2-acetaminofluorene. L02 human hepatocytes can survive for 10 wk after transplantation and express human albumin for 8 wk. L02 human hepatocytes can proliferate and express human albumin for 6 mo after transplantation to the rats treated with Retrorsine. The chimeric L02 human hepatocytes, which then underwent transplantation into tolerant rats, were normal in morphogenesis, biochemistry and function.
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Laconi E, Doratiotto S, Vineis P. The microenvironments of multistage carcinogenesis. Semin Cancer Biol 2008; 18:322-9. [PMID: 18456510 DOI: 10.1016/j.semcancer.2008.03.019] [Citation(s) in RCA: 37] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/20/2008] [Accepted: 03/25/2008] [Indexed: 12/19/2022]
Abstract
Overt neoplasia is often the result of a chronic disease process encompassing an extended segment of the lifespan of any species. A common pathway in the natural history of the disease is the appearance of focal proliferative lesions that are known to act as precursors for cancer development. It is becoming increasingly apparent that the emergence of such lesions is not a cell-autonomous phenomenon, but is heavily dependent on microenvironmental cues derived from the surrounding tissue. Specific alterations in the tissue microenvironment that can foster the selective growth of focal lesions are discussed herein. Furthermore, we argue that a fundamental property of focal lesions as it relates to their precancerous nature lies in their altered growth pattern as compared to the tissue where they reside. The resulting altered tissue architecture translates into the emergence of a unique tumor microenvironment inside these lesions, associated with altered blood vessels and/or blood supply which in turn can trigger biochemical and metabolic changes fueling tumor progression. A deeper understanding of the role(s) of tissue and tumor microenvironments in the pathogenesis of cancer is essential to design more effective strategies for the management of this disease.
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Affiliation(s)
- Ezio Laconi
- Dipartimento di Scienze e Tecnologie Biomediche, Sezione di Patologia Sperimentale, Università di Cagliari, 09125 Cagliari, Italy.
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Alwayn IPJ, Verbesey JE, Kim S, Roy R, Arsenault DA, Greene AK, Novak K, Laforme A, Lee S, Moses MA, Puder M. A critical role for matrix metalloproteinases in liver regeneration. J Surg Res 2008; 145:192-8. [PMID: 18222481 DOI: 10.1016/j.jss.2007.04.002] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/30/2006] [Revised: 03/01/2007] [Accepted: 04/01/2007] [Indexed: 12/16/2022]
Abstract
BACKGROUND Matrix metalloproteinases (MMPs), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) are mediators of liver regeneration. To determine whether MMPs are required for normal hepatic regeneration, we performed 67% hepatectomies on mice treated with a broad-spectrum MMP-inhibitor, and assessed the effect on liver regeneration and urinary MMP activity. METHODS Mice were subjected to sham operations, 67% hepatectomy, or 67% hepatectomy plus treatment with the broad-spectrum MMP inhibitor Marimastat. Urine collected preoperatively and for 8 d postoperatively was tested for MMP-2 and MMP-9 activity using zymography. Serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, bilirubin, TNF-alpha, IL-6, and hepatocyte growth factor levels were measured. Liver sections were analyzed by CD31 immunohistochemistry and microvessel density. Mitotic index and proliferating cell nuclear antigen labeling index were determined. RESULTS The mean regenerating liver weight on postoperative day 8 was 0.72 +/- 0.01 grams for the hepatectomy Marimastat group, and 0.83 +/- 0.02 grams for the hepatectomy control group (P < 0.001). Urinary MMP-9 activity was elevated during hepatic regeneration, and decreased on postoperative day 8 when the liver returned to its preoperative mass. In contrast, urine from hepatectomy Marimastat mice, in which liver regeneration was successfully inhibited, showed consistently low levels of MMP-2 and MMP-9 activity. The hepatectomy Marimastat group also exhibited elevated serum IL-6 levels on post-operative day 8, while serum TNF-alpha soluble receptor II levels were unchanged. Hepatocyte growth factor levels were not significantly different between the control hepatectomy and hepatectomy Marimastat groups at days 2, 4, and 8. Liver microvessel density was reduced in the hepatectomy Marimastat group at day 4. Mitotic index and proliferating cell nuclear antigen index were significantly decreased in the Marimastat hepatectomy group at post-operative day 2. CONCLUSIONS The broad-spectrum MMP-inhibitor Marimastat inhibits liver regeneration. Microvessel density is reduced at day 4. Furthermore, urinary MMP-9 is elevated during liver regeneration, and this effect is not observed when regeneration is inhibited by the broad-spectrum MMP-inhibitor Marimastat.
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Affiliation(s)
- Ian P J Alwayn
- Department of Surgery and The Vascular Biology Program, Children's Hospital, Boston, Massachusetts 02115, USA
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Bird TG, Lorenzini S, Forbes SJ. Activation of stem cells in hepatic diseases. Cell Tissue Res 2008; 331:283-300. [PMID: 18046579 PMCID: PMC3034134 DOI: 10.1007/s00441-007-0542-z] [Citation(s) in RCA: 124] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/01/2007] [Accepted: 10/23/2007] [Indexed: 02/06/2023]
Abstract
The liver has enormous regenerative capacity. Following acute liver injury, hepatocyte division regenerates the parenchyma but, if this capacity is overwhelmed during massive or chronic liver injury, the intrinsic hepatic progenitor cells (HPCs) termed oval cells are activated. These HPCs are bipotential and can regenerate both biliary epithelia and hepatocytes. Multiple signalling pathways contribute to the complex mechanism controlling the behaviour of the HPCs. These signals are delivered primarily by the surrounding microenvironment. During liver disease, stem cells extrinsic to the liver are activated and bone-marrow-derived cells play a role in the generation of fibrosis during liver injury and its resolution. Here, we review our current understanding of the role of stem cells during liver disease and their mechanisms of activation.
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Affiliation(s)
- T G Bird
- MRC/University of Edinburgh Centre for Inflammation Research, The Queen's Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh, EH16 4TJ, UK.
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Improvement of the survival rate by fetal liver cell transplantation in a mice lethal liver failure model. Transplantation 2007; 84:1233-9. [PMID: 18049107 DOI: 10.1097/01.tp.0000287967.54222.4d] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
Abstract
BACKGROUND The use of cell transplantation as an alternative therapy for orthotopic liver transplantation has been widely anticipated due to a chronic donor shortage. We previously reported the method used to enrich hepatic progenitor cells (HPCs) forming cell aggregations. In this study, we transplanted HPCs into the liver injury model mice to determine whether HPC transplantation may improve the liver dysfunction. METHODS We obtained donor cells from E13.5 fetal livers of green fluorescent protein (GFP) transgenic mice. We transplanted GFP-positive fetal liver cells into the transgenic mice which express diphtheria toxin (DT) receptors under the control of an albumin enhancer/promoter. Subsequently, we induced selective liver injury to recipient mice by DT administration. We then evaluated the engraftment of the transplanted cells and their effect on survivorship. RESULTS The low dose of DT induced sublethal liver injury and the high dose of DT was lethal to the liver injury model mice. The transplanted GFP-positive cells were engrafted into the recipient livers and expressed albumin, resembling mature hepatocytes. They continued to proliferate, forming clusters. The survival rate at 25 days after transplantation of the cell-transplanted group (8 of 20; 40.0%) was improved significantly (P=0.0047) in comparison to that of the sham-operated group (0 of 20; 0%). CONCLUSIONS The transplanted cells were engrafted and repopulated the liver of recipient mice, resulting in the improvement of the survival rate of the liver injury model mice. We therefore propose that HPCs are a desirable cell source for cell transplantation.
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Best DH, Coleman WB. Bile duct destruction by 4,4'-diaminodiphenylmethane does not block the small hepatocyte-like progenitor cell response in retrorsine-exposed rats. Hepatology 2007; 46:1611-9. [PMID: 17705295 DOI: 10.1002/hep.21876] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/24/2023]
Abstract
UNLABELLED Liver regeneration after surgical partial hepatectomy (PH) in retrorsine-exposed rats is accomplished through the outgrowth and expansion of small hepatocyte-like progenitor cells (SHPCs). The cells of origin for SHPCs and their tissue niche have not been identified. Nevertheless, some investigators have suggested that SHPCs may represent an intermediate or transitional cell type between oval cells and mature hepatocytes, rather than a distinct progenitor cell population. We investigated this possibility through the targeted elimination of oval cell proliferation secondary to bile duct destruction in retrorsine-exposed rats treated with 4,4'-diaminodiphenylmethane (DAPM). Fischer 344 rats were treated with 2 doses (30 mg/kg body weight) retrorsine (at 6 and 8 weeks of age) followed by PH 5 weeks later. Twenty-four hours before PH, select animals were given a single dose of DAPM (50 mg/kg). Treatment of rats with DAPM produced severe bile duct damage but did not block liver regeneration. Oval cells were never seen in the livers of DAPM-treated retrorsine-exposed rats after PH. Rather, liver regeneration in these rats was mediated by the proliferation of SHPCs, and the cellular response was indistinguishable from that observed in retrorsine-exposed rats after PH. SHPC clusters emerge 1 to 3 days post-PH, expand through 21 days post-PH, with normalization of the liver occurring by the end of the experimental interval. CONCLUSION These results provide direct evidence that SHPC-mediated liver regeneration does not require oval cell activation or proliferation. In addition, these results provide strong evidence that SHPCs are not the progeny of oval cells but represent a distinct population of liver progenitor cells.
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Affiliation(s)
- D Hunter Best
- Department of Pathology and Laboratory Medicine, Curriculum in Toxicology, UNC Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC, USA
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Khuu DN, Najimi M, Sokal EM. Epithelial cells with hepatobiliary phenotype: is it another stem cell candidate for healthy adult human liver? World J Gastroenterol 2007; 13:1554-60. [PMID: 17461448 PMCID: PMC4146898 DOI: 10.3748/wjg.v13.i10.1154] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM To investigate the presence and role of liver epithelial cells in the healthy human adult liver. METHODS Fifteen days after human hepatocyte primary culture, epithelial like cells emerged and started proliferating. Cell colonies were isolated and subcultured for more than 160 d under specific culture conditions. Cells were analyzed for each passage using immunofluorescence, flow cytometry and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS Flow cytometry analysis demonstrated that liver epithelial cells expressed common markers for hepatic and stem cells such as CD90, CD44 and CD29 but were negative for CD34 and CD117. Using immunofluorescence we demonstrated that liver epithelial cells expressed not only immature (alpha-fetoprotein) but also differentiated hepatocyte (albumin and CK-18) and biliary markers (CK-7 and 19), whereas they were negative for OV-6. RT-PCR analysis confirmed immunofluorescence data and revealed that liver epithelial cells did not express mature hepatocyte markers such as CYP2B6, CYP3A4 and tyrosine amino-transferase. Purified liver epithelial cells were transplanted into SCID mice. One month after transplantation, albumin positive cell foci were detected in the recipient mouse parenchyma. CONCLUSION According to their immature and bipotential phenotype, liver epithelial cells might represent a pool of precursors in the healthy human adult liver other than oval cells.
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Affiliation(s)
- Dung-Ngoc Khuu
- Université Catholique de Louvain, Laboratory of Pediatric Hepatology and Cell Therapy, Brussels, Belgium
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50
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Best DH, Coleman WB. Treatment with 2-AAF blocks the small hepatocyte-like progenitor cell response in retrorsine-exposed rats. J Hepatol 2007; 46:1055-63. [PMID: 17434228 PMCID: PMC1974854 DOI: 10.1016/j.jhep.2007.01.040] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/12/2006] [Revised: 01/05/2007] [Accepted: 01/09/2007] [Indexed: 12/04/2022]
Abstract
BACKGROUND/AIMS Liver regeneration after partial hepatectomy (PH) in retrorsine-exposed rats is accomplished through proliferation and differentiation of small hepatocyte-like progenitor cells (SHPCs). The cells of origin of SHPCs are not known. We investigated the possibility that SHPCs are directly derived from oval cells, a known liver progenitor cell, by combining the retrorsine/PH (RP) model with 2-acetamidofluorene (2-AAF), an anti-mitotic agent that elicits an oval cell reaction in response to liver deficit. METHODS Male Fischer 344 rats were treated with retrorsine (30 mg/kg ip) at 6 and 8 weeks of age, with PH 5 weeks after the final treatment. Seven days prior to PH, a 21-day 2-AAF (50mg) time-release pellet was inserted subcutaneously. Livers were harvested at 3, 7, 10, 14, and 21-days post-PH. RESULTS Liver sections from animals treated with 2-AAF/retrorsine/PH (2-AAF/RP) contain significant numbers of proliferating oval cells, but no SHPCs at 7-days post-PH, while RP animals exhibit significant numbers of SHPCs and minimal oval cell reaction. Between 10 and 14-days post-PH, new hepatocyte clusters appear in 2-AAF/RP treated rats. Labeling of proliferating oval cells with BrdU at 6-days post-PH demonstrated that these new hepatocytes represent the progeny of differentiating oval cells. CONCLUSIONS The observed differences in progenitor cell responses between 2-AAF/RP and RP animals strongly suggest that SHPCs are not the progeny of oval cell precursors, but represent an independent liver progenitor cell population.
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Affiliation(s)
- D Hunter Best
- Department of Pathology and Laboratory Medicine, Curriculum in Toxicology, UNC Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA
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