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Comment on “ Novel therapeutic diiminoquinone exhibits anticancer effects on human colorectal cancer cells in two-dimensional and three-dimensional in vitro models” Chaolin Deng1, Bin Wu1 1 Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China. Corresponding author: Bin Wu, MD, Chief Doctor, Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, No. 1 Shuaifuyuan Road, Wangfujing, Dongcheng District, Beijing 100730, China. Email:wubin0279@hotmail.com Author contributions: Author contributions: Wu B designed and revised the manuscript; Deng CL wrote the manuscript; Abstract We recently read with interest the article“ Novel therapeutic diiminoquinone exhibits anticancer effects on human colorectal cancer cells in two-dimensional and three-dimensional in vitro models”, this original article was using 2D/3D models of CRC demonstrating the efficacy of anti-tumor with Diiminoquinone (DQI). through further research and analysis, we feel obliged to offer some relevant insights. These comments may deepen our understanding in the context of the three-dimensional (3D) drug screening models and pathological subtypes have different responses to chemotherapies. Keywords: Colorectal cancer, Diiminoquinone, 3D-bioprinting, Pathological subtypes, Drug screening Core Tip: Colorectal cancer is one of the most prevalent cancers worldwide, for inoperable colorectal cancer, chemotherapy is the main treatment option. We recently read with interest the article” Novel therapeutic diiminoquinone exhibits anticancer effects on human colorectal cancer cells in two-dimensional and three-dimensional in vitro models”. We would like to share our opinion on this article and hope that it will be of some help to the authors and readers   TO THE EDITOR We read with pleasure the article by Alissar Monzer et al. Within this article, the authors assessed the potential anti-tumor effects of Diiminoquinone（DIQ）in Colorectal cancer(CRC) two-dimensional (2D) or three-dimensional (3D) models. Furthermore, the in vivo anti-tumor ability assessment was evaluated using an in vivo model. The authors showed that DIQ could potentially be therapeutic, and inhibit proliferation, invasion, and migration of the tumor cells, by Wnt/-catenin and AKT and ERK pathways. The salient highlight of this article was using multiple 3D models to independent validation of this compound for treatment in CRC. The tumor microenvironment provides essential signals to guide tumor growth and survival[1, 2]. The conventional 2D culture system has long been used in studies of chemotherapy responses. However, the 2D model cannot maintain the interactions of tumor cells, which further complicates the successful translation into the clinic. The authors successfully established the Sphere formation with tumor cell lines, the salient feature of Cancer stem cells (CSCs), and derived-organoid culture with colon tumor tissue. In subsequent studies, DIQ triggered selective tumor inhibition, but not of normal cells, which verified the safety and efficacy. We found some details through further research and analysis and feel obliged to offer some relevant insights. The main role of 3D culture models aimed to mimic the native tumor microenvironment. Tumor microenvironment (TME) and intercellular communication can affect various signal transduction processes in cancer cells. The sphere formation assay is a classic functional approach for studying tumor stem cells. However, this method lack cell-to-cell communication in cancer cells and interstitial cells. 3D-printing is capable of fabricating complex 3D structures with multiple cell types, extracellular matrix components, and growth and signaling factors[3], we consider 3D-bioprinting can use multiple cell types in 3D architecture to better imitate the TME and then in developing more accurate therapeutic drug compared to 2D models[4]. Recently, various 3D-bioprinting drug screening models were reported to achieve higher predictive rates for drug response and novel drug development[2, 5]. Furthermore, 3D-bioprinting could as the foundation for future precision medicine for drug screening. Another interesting finding by Alissar Monzer et al was the DIQ showed the chemotherapy sensitivity in mucinous neoplasms, special pathological subtypes in CRC[6]. The first‐line treatment in patients with metastatic colorectal cancer (mCRC) usually involves chemotherapy. Previous studies demonstrated that patients with mucinous tumors with lesser involves either irinotecan‐ or oxaliplatin‐based chemotherapy benefit with than patients with non-mucinous tumor[7]. In the following, the anti-tumor mechanism of DQI was proved that was activating γH2AX expression to inducer of DNA damage. However, due to the small size of the tissue sample and the low rate of deriving colon patient-derived organoids, the effectiveness of chemotherapy in mucinous tumors needs to be addressed with further research. This may provide directions for targeted therapy with differentiated subtype interventions. In summary, this original article was using 2D/3D models of CRC demonstrating the efficacy of anti-tumor with DQI. However, in light of the complexity of CRC tumor and individual differences in patients, the detailed underlying mechanisms still need much more studies for rational and effective treatment innovation.  1 Habanjar O, Diab-Assaf M, Caldefie-Chezet F, Delort L. 3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages. Int J Mol Sci 2021; 22(22) [PMID: 34830082 PMCID: PMC8618305 DOI: 10.3390/ijms222212200] 2 Ramzy GM, Koessler T, Ducrey E, McKee T, Ris F, Buchs N, Rubbia-Brandt L, Dietrich PY, Nowak-Sliwinska P. Patient-Derived In Vitro Models for Drug Discovery in Colorectal Carcinoma. Cancers (Basel) 2020; 12(6) [PMID: 32486365 PMCID: PMC7352800 DOI: 10.3390/cancers12061423] 3 Sbirkov Y, Molander D, Milet C, Bodurov I, Atanasov B, Penkov R, Belev N, Forraz N, McGuckin C, Sarafian V. A Colorectal Cancer 3D Bioprinting Workflow as a Platform for Disease Modeling and Chemotherapeutic Screening. Front Bioeng Biotechnol 2021; 9: 755563 [PMID: 34869264 PMCID: PMC8638705 DOI: 10.3389/fbioe.2021.755563] 4 Neufeld L, Yeini E, Reisman N, Shtilerman Y, Ben-Shushan D, Pozzi S, Madi A, Tiram G, Eldar-Boock A, Ferber S, Grossman R, Ram Z, Satchi-Fainaro R. Microengineered perfusable 3D-bioprinted glioblastoma model for in vivo mimicry of tumor microenvironment. Sci Adv 2021; 7(34) [PMID: 34407932 PMCID: PMC8373143 DOI: 10.1126/sciadv.abi9119] 5 Ma K, Zhao T, Yang L, Wang P, Jin J, Teng H, Xia D, Zhu L, Li L, Jiang Q, Wang X. Application of robotic-assisted in situ 3D printing in cartilage regeneration with HAMA hydrogel: An in vivo study. J Adv Res 2020; 23: 123-132 [PMID: 32099674 PMCID: PMC7030996 DOI: 10.1016/j.jare.2020.01.010] 6 Taieb J, Shi Q, Pederson L, Alberts S, Wolmark N, Van Cutsem E, de Gramont A, Kerr R, Grothey A, Lonardi S, Yoshino T, Yothers G, Sinicrope FA, Zaanan A, Andre T. Prognosis of microsatellite instability and/or mismatch repair deficiency stage III colon cancer patients after disease recurrence following adjuvant treatment: results of an ACCENT pooled analysis of seven studies. Ann Oncol 2019; 30(9): 1466-1471 [PMID: 31268130 PMCID: PMC7360150 DOI: 10.1093/annonc/mdz208] 7 Kwon M, Rubio G, Nolan N, Auteri P, Volmar JA, Adem A, Javidian P, Zhou Z, Verzi MP, Pine SR, Libutti SK. FILIP1L Loss Is a Driver of Aggressive Mucinous Colorectal Adenocarcinoma and Mediates Cytokinesis Defects through PFDN1. Cancer Res 2021; 81(21): 5523-5539 [PMID: 34417201 PMCID: PMC8563430 DOI: 10.1158/0008-5472.CAN-21-0897]

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