Use of zebrafish embryos as avatar of patients with pancreatic cancer: A new xenotransplantation model towards personalized medicine

Table 1 Patients characteristics (n = 15)

Characteristics
Mean age, years ± SD	71.2 ± 9.9 (44.1-83.2)
M:F, n (%)	8:7 (53.3:46.7)
Mean BMI, kg/m2 ± SD	26.4 ± 5.1 (17.6-40.4)
Ca 19.9, U/mL ± SD	347.1 ± 543.0 (1.7-2094)
Type of surgical procedure, n (%)
Pancreatoduodenectomy	11 (73.3)
Distal splenopancreatectomy	3 (20)
Total splenopancreatectomy	1 (6.7)
Vascular resection, n (%)	4 (26.7)
Grade of differentiation, n (%)
G2/3	11 (73.3)
G3/3	4 (26.7)
Mean tumour dimension, cm	3.3 ± 1.2 (1.5-5.0)
Mean harvest lymph nodes, n	40.6 ± 18.0 (19-74)
Mean positive lymph nodes, n	5.7 ± 6.0 (0-22)
T status, n (%)
T1	2 (13.3)
T2	7 (46.7)
T3	6 (40)
N status, n (%)
N0	1 (6.6)
N1	7 (46.7)
N2	7 (46.7)
Stage, n (%)
IB	1 (6.7)
IIB	6 (40)
III	8 (53.3)
Angioinvasion, n (%)	2 (13.3)
Perineural infiltration, n (%)	11 (73.4)
Vascular infiltration, n (%)	2 (13.3)

BMI: Body mass index; M: Male; F: Female.

Table 2 Survival rate of the xenotransplanted zebrafish embryos

No. of case	1 d post injection					2 d post injection
	Control group	GEM	GEMO-X	GEM/ nab-P	FOLF-OXIRI	Control group	GEM	GEMOX	GEM/nab-P
1	0.909	0.833	0.917	0.917	0.900	0.818	0.667	0.833	0.583
2	0.417	0.800	0.917	0.667	0.833	0.333	0.583	0.667	0.500
3	0.833	1.000	0.636	0.917	0.833	0.417	0.818	0.182	0.75
4	0.682	0.438	0.7692	0.750	0.647	0.545	0.375	0.692	0.688
5	0.533	0.750	0.667	1.000	0.555	0.533	0.625	0.267	0.824
6	0.476	0.429	0.462	0.563	0.688	0.477	0.429	0.462	0.563
7	0.857	0.692	0.846	0.733	0.688	0.667	0.538	0.615	0.267
8	0.864	0.533	0.765	0.933	0.588	0.727	0.533	0.539	0.800
9	0.652	0.471	0.058	0.471	0.556	0.609	0.353	0.059	0.471
10	0.800	0.611	0.733	0.800	0.800	0.150	0.5	0.667	0.600
11	0.917	0.923	1.000	0.800	1.000	0.500	0.462	0.571	0.500
12	0.789	0.75	0.687	0.556	0.529	0.632	0.625	0.312	0.167
13	0.957	0.824	0.75	0.765	0.833	0.739	0.647	0.500	0.471
14	0.700	0.467	0.556	0.500	0.333	0.550	0.267	0.278	0.167
15	0.846	0.786	0.786	0.538	0.833	0.615	0.357	0.429	0.462
Mean survival rate	0.749	0.687	0.703	0.727	0.708	0.554	0.519	0.471	0.521

GEM: Gemcitabine; GEMOX: Gemcitabine + Oxaliplatin; GEM/nab-P: Gemcitabine + nab-Paclitaxel; FOLFOXIRI: 5-Fluorouracil + Folinic acid + Oxaliplatin + Irinotecan.

Table 3 Results of exposure to chemotherapy in xenotransplanted zebrafish embryos

Mean relative tumor area					Mean difference				95%CI				P value
Control group	FOLFOXIRI	GEMOX	GEM/nab-P	GEM	k vs FOLFOXIRI	k vs GEMOX	k vs GEM/nab-P	k vs Gemcitabine	k vs FOLFOXIRI	k vs GEMOX	k vs GEM/nab-P	k vs Gemcitabine	k vs FOLFOXIRI	k vs GEMOX	k vs GEM/nab-P	k vs Gemcitabine
3.026	0.790	1.243	0.988	1.213	2.236	1.784	2.038	1.813	0.286 to 4.187	-0.167 to 3.734	-0.047 to 4.123	-0.198 to 3.823	0.020	0.081	0.057	0.087
1.964	0.755	1.188	0.529	1.674	1.209	0.776	1.435	0.290	-0.016 to 2.434	-0.522 to 2.073	0.099 to 2.771	-0.978 to 1.558	0.054	0.361	0.032	0.936
1.699	0.524	1.599	1.238	0.706	1.175	0.100	0.461	0.993	-0.026 to 2.376	-1.348 to 1.548	-0.645 to 1.567	-0.113 to 2.099	0.057	0.999	0.659	0.088
1.604	0.951	1.990	1.767	1.013	0.653	-0.385	-0.163	0.592	-0.452 to 1.759	-1.551 to 0.780	-1.269 to 0.942	-0.724 to 1.907	0.381	0.817	0.987	0.616
2.357	2.260	1.689	1.566	1.263	0.097	0.668	0.792	1.094	-2.112 to 0.261	-1.363 to 2.700	-0.679 to 2.262	-0.479 to 2.668	> 0.990	0.817	0.458	0.244
2.098	0.714	1.423	1.130	1.672	1.384	0.674	0.968	0.426	-2.112 to 0.261	-0.826 to 2.175	-0.411 to 2.346	-1.075 to 1.926	0.034	0.610	0.236	0.879
2.170	1.663	1.746	1.205	0.965	0.507	0.424	0.966	1.205	-2.112 to 0.261	-0.760 to 1.607	-0.478 to 2.409	0.022 to 2.388	0.560	0.770	0.270	0.040
1.224	0.489	1.802	1.189	1.221	0.735	-0.578	0.035	0.003	-2.112 to 0.261	-2.130 to 0.975	-1.311 to 1.382	-1.549 to 1.556	0.730	0.750	> 0.990	> 0.990
1.606	0.710	0.423	0.606	0.945	0.896	1.183	1.000	0.662	-2.112 to 0.261	-0.383 to 2.748	0.083 to 1.918	-0.417 to 1.740	0.057	0.183	0.029	0.342
1.046	0.575	1.037	0.468	0.514	0.471	0.009	0.579	0.533	-2.112 to 0.261	-1.094 to 1.113	-0.557 to 1.714	-0.586 to 1.650	0.643	> 0.990	0.506	0.563
2.669	1.203	1.207	1.428	1.371	1.466	1.462	1.241	1.298	-2.112 to 0.261	-1.407 to 4.331	-1.326 to 3.807	-1.268 to 3.864	0.338	0.492	0.537	0.499
1.371	0.852	0.705	1.383	1.077	0.520	0.666	-0.012	0.294	-0.667 to 1.706	-0.775 to 2.107	-1.452 to 1.429	-0.774 to 1.363	0.627	0.584	> 0.990	0.888
1.648	0.8481	0.8028	1.174	1.273	0.7996	0.8449	0.4737	0.3748	0.019 to 1.580	-0.030 to 1.720	-0.3717 to 1.319	-0.471 to 1.220	0.043	0.061	0.433	0.632
1.275	1.704	0.888	2.157	1.683	-0429	0.387	-0.882	-0.408	-1.975 to 1.117	-0.957 to 1.731	-2.428 to 0.664	-1.954 to 1.138	0.879	0.864	0.394	0.896
1.041	1.966	1.167	0.789	1.091	-0.926	-0.126	0.252	-0.050	-2.112 to 0.261	-1.356 to 1.104	-1.035 to 1.539	-1.281 to 1.180	0.162	> 0.990	0.963	> 0.990

k: Control group; GEM: Gemcitabine; GEMOX: Gemcitabine + Oxaliplatin; GEM/nab-P: Gemcitabine + nab-Paclitaxel; FOLFOXIRI: 5-Fluorouracil + Folinic acid + Oxaliplatin + Irinotecan.

Table 4 Percentage reduction of the mean relative tumor area

No. of case	k vs FOLFOXIRI (%)	k vs GEMOX (%)	k vs Gem/nab-P (%)	k vs Gemcitabine (%)
1	-74	-59	-67	-60
2	-62	-40	-73	-15
3	-69	-6	-27	-58
4	-41	24	10	-37
5	-4	-28	-34	-46
6	-66	-32	-46	-20
7	-23	-20	-44	-56
8	-60	47	-3	0
9	-56	-74	-62	-41
10	-45	-1	-55	-51
11	-55	-55	-46	-49
12	-38	-49	1	-21
13	-49	-51	-29	-23
14	34	-30	69	32
15	54	-8	-38	-14

k: Control group; GEM: Gemcitabine; GEMOX: Gemcitabine + Oxaliplatin; GEM/nab-P: Gemcitabine + nab-Paclitaxel; FOLFOXIRI: 5-Fluorouracil + Folinic acid + Oxaliplatin + Irinotecan.

Table 5 Analysis of the percentage reduction of the mean relative tumor area in the treated subgroups in comparison to the control group

Threshold value	Number of cases, n (%)
	k vs FOLFOXIRI	k vs GEMOX	k vs Gem/nab-P	k vs Gemcitabine
-10%	13 (87)	13 (87)	12 (80)	13 (87)
-20%	12 (80)	10 (67)	11 (73)	11 (73)
-30%	11 (73)	8 (53)	9 (60)	8 (53)
-40%	10 (67)	6 (40)	7 (47)	7 (47)
-50%	7 (47)	5 (33)	4 (27)	4 (27)
-60%	5 (33)	1 (7)	4 (27)	1 (7)
-70%	1 (7)	1 (7)	1 (7)	0 (0)
-80%	0 (0)	0 (0)	0 (0)	0 (0)

k: Control group; GEM: Gemcitabine; GEMOX: Gemcitabine + Oxaliplatin; GEM/nab-P: Gemcitabine + nab-Paclitaxel; FOLFOXIRI: 5-Fluorouracil + Folinic acid + Oxaliplatin + Irinotecan.

Table 6 Calculation of the conversion factor for the test with xenotransplanted zebrafish embryos

	Study	Percentage of partial response reported in literature	Conversion Factor for xenotransplanted zebrafish embryos	Mean conversion factor (P = 0.626)
k vs FOLFOXIRI	Conroy et al[7], 2011	31	-60.1	-60.4
	Tong et al[40], 2018	35.9	-56.2
	Lakatos et al[41], 2017	18.8	-69.8
	Nitsche et al[42], 2015	42.8	-50.7
	Sadot et al[43], 2015	29	-61.7
	Wang et al[44], 2019	32.3	-59.1
k vs GEMOX	Shi et al[46], 2007	26.8	-55.2	-59.3
	Li et al[47], 2010	25	-56.8
k vs Gem/nab-P	Von Hoff et al[48], 2013	23	-61.3	-62.4
	Casanova-Martinez et al[49], 2018	20	-64.0
	Karasic et al[50], 2019	21.1	-63.0
	Montes et al[51], 2017	23	-61.3
k vs Gemcitabine	Conroy et al[7], 2011	16	-63.9	-63.7
	Louvet et al[52], 2005	17.3	-62.8
	Ergun et al[53], 2018	15.3	-64.5

k: Control group; GEM: Gemcitabine; GEMOX: Gemcitabine + Oxaliplatin; GEM/nab-P: Gemcitabine + nab-Paclitaxel; FOLFOXIRI: 5-Fluorouracil + Folinic acid + Oxaliplatin + Irinotecan.