Isogenic human cell lines comprising at least one mutated cancer allele under the control of the cell line endogenous promoter, which corresponds to the wild-type cancer allele promoter are disclosed, as well as an in vitro process for determining sensitivity/resistance of a patient suffering from a tumor to a pharmacological agent comprising the following steps: a) identifying at least one mutated cancer allele in a tissue affected by a tumor of said patient; b) providing an isogenic human cell line representative of the tissue, wherein the cell line comprises at least the identified mutated cancer allele, which is under the control of the cell line endogenous promoter corresponding to the wild-type cancer allele promoter; c) putting in contact said cell line with the pharmacological agent; d) determining a variation of proliferation, apoptosis or cytotoxicity of the cell line in presence of the pharmacological agent; wherein the variation of proliferation, apoptosis or cytotoxicity is indicative of the sensitivity/resistance of the patient tumor to the pharmacological agent. Claims 1. Isogenic human cell line, characterized in that said cell line comprises at least one mutated cancer allele, in that said at least one mutated cancer allele is under the control of an endogenous promoter of said cell line, said endogenous promoter being corresponding to the wild-type cancer allele promoter, and in that said cancer allele is selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4, FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (pl6) , BRCAl, BRCA2 , PTCHl, VHL, SMAD4, PERl, MENl, NFl, NF2, ATM, PTPRD. 2. Isogenic human cell line according to claim 1, wherein said cell line carries at least two mutated cancer alleles, wherein said at least two cancer alleles are selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3 , ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4, FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (plβ) , BRCAl, BRCA2 , PTCHl, VHL, SMAD4, PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 3. Isogenic human cell line according to claim 1 or claim 2, wherein said at least one mutated cancer allele is selelected among the mutated cancer alleles listed in Table 2a. 4. Isogenic human cell line according to any one of claims 1 to 3, wherein said human cell line is selected among MCFlOA, hTERT-HMEl, HTERT-RPE-I, HCT 116, DLD-I, SW48, NuLi, CuFi, CHON-001, CHON-002, BJ-5ta, hTERT-HMEl (ME16C) , hTERT RPE-I, hTERT-HPNE, NeHepLxHT, T HESCs, RWPE-I, RWPE-2, WPE-stem, WPE-int, WPE1-NA22, WPE1-NB14, WPEl-NBIl, WPEl-NB26, RWPE2-W99, WPMY-I, WPE1-NB26-64 , WPE1-NB26-65, HBE4-E6/E7 [NBE4-E6/E7] , JVM-13, MeT-5A, BBM, BZR, BEAS-2B, MCF 1OA, MCF 1OF, MCF-10-2A, B-3, HBE4-E6/E7-C1 , HK-2, CHON-001, CHON-002, HS-5, PWR-IE, THLE-3, HCE-2 [50. Bl], 46BR. IN, BRISTOL 8, AGLCL, C211, GM1899A, GS-109-V-63, GS-109-V-34, H9, HFFF2, HFLl, HG261, HH-8, HL, Hs 68, Hs 888Lu, HsI. Tes, IM 9, MRC-5 pdl9, MRC-5 pd25, MRC-5 pd30, MRC-5 pd30, MRC-5 SVl TGl, MRC-5 SVl TG2, MRC-5 SV2 , MRC-7, MRC-9, MT-2, PNTlA, PNTlA (SERUM FREE), PNT2, PNT2 (SERUM FREE) , SVCT, SVCT-MI2, TK6, TK6TGR, TOU (TOU 1-2), WI 26 VA4 , WI 38, WI 38VA13 Subline 2RA, WiDr, WIL2 NS, WIL2.NS.6TG, WILCL, OVCAR-5, OVCAR-4 , OVCAR-3, NCI-H522, NCI-H460, NCI-H322M, NCI-H23, NCI-H226, NCI/ADR-RES, MOLT-4, MDA-N, MDA-MB-435, MDA-MB-231, MCF7, Malme-3M, M14, LOXIMVl, KM12, K-562, IGROVl, HT-29, Hs 578T, HOP-92, HOP-62, HL-60, HCT-15, HCT-116, HCC-2998, EKVX, DU-145, COLO-205, CCRF-CEM, CAKI-I, BT-549, ACHN, A549, A498, 786-0 cell lines. 5. Isogenic human cell line according to any one of the preceding claims, wherein the mutated BRAF cancer allele carries the mutation V600E as shown in SEQ ID No . : 3. 6. Isogenic human cell line according to any one of the preceding claims, wherein the mutated EGFR cancer allele carries the mutation delE746-A750 as shown in SEQ ID No . : 5. 7. Isogenic human cell line according to any one of the preceding claims, wherein the mutated PIK3CA cancer allele carries the mutations E545K and H1047R as shown in SEQ ID No . : 1 and 2, respectively. 8. Isogenic human cell line according to any one of the preceding claims, wherein the mutated CTNNBl cancer allele carries the mutation T41A as shown in SEQ ID No . : 6. 9. Isogenic human cell line according to any one of the preceding claims, wherein the mutated PTEN cancer allele carries the mutation R130* as shown in SEQ ID No . : 7. 10. Isogenic human cell line according to any one of the preceding claims, wherein said cell line carries at least one detectable marker. 11. Isogenic human cell line according to claim 10, wherein said at least one marker is selected among a fluorescent, radioactive, luminescent, phosphorescent marker. 12. Isogenic human cell line according to any one of the preceding claims, wherein said cell line carries at least one knocked-out or inactivated tumor suppressor gene. 13. Isogenic human cell line according to claim 12, wherein said at least one tumor suppressor gene is selected among PTEN, TP53, APC, p21, RbI, BUBl, BRCAl, BRCA2, PTCH, VHL, SMAD4, PERl, TSC2, CDKN2A, DCC, MEN-I, NFl, ATM, PTPRD, LRPlB and NF2. 14. Use of an isogenic human cell line according to any one of claims 1 to 13 for generating xenografts apt to induce tumor growth in a non-human laboratory animal model. 15. Use of an isogenic human cell line according to any one of claims 1 to 13 for producing non-human transgenic laboratory animals susceptible to develop a tumor, said tumor carrying at least one mutated cancer allele, said cancer allele being selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4, FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (plβ) , BRCAl, BRCA2, PTCHl, VHL, SMAD4, PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 16. Use according to claim 15, wherein said non-human transgenic laboratory animals carrying a tumor are used for determining the sensitivity/resistance of said tumor to a pharmacological agent administered to said transgenic animals . 17. In vitro process for determining sensitivity/resistance of a patient suffering from a tumor to a pharmacological agent, characterized in that said process comprises: a) identifying at least one mutated cancer allele in a tissue affected by a tumor of said patient; b) providing an isogenic human cell line representative of said tissue, said cell line comprising at least said mutated cancer allele, wherein said cancer allele is under the control of an endogenous promoter of said cell line, said endogenous promoter being corresponding to the wild-type cancer allele promoter; c) putting in contact said isogenic cell line with said pharmacological agent; d) determining a variation of proliferation, cytotoxicity and/or apoptosis of said isogenic cell line in presence of said pharmacological agent; said variation of proliferation, cytotoxicity and/or apoptosis being indicative of said sensitivity/resistance of said patient to said pharmacological agent. 18. Process according to claim 17, wherein said process further comprises: bl) providing a wild-type isogenic human cell line representative of said tissue, being said wild-type isogenic human cell line free of said mutated cancer allele; cl) putting in contact said wild-type isogenic cell line with said pharmacological agent; dl) determining a variation of proliferation, cytotoxicity and/or apoptosis of said wild-type isogenic cell line in presence of said pharmacological agent. 19. Process according to claim 17 and claim 18, wherein said sensitivity/resistance is evaluated as the relative variation of proliferation, apoptosis and/or cytotoxicity between said isogenic human cell line comprising said at least mutated cancer allele and said wild-type isogenic human cell line. 20. Process according to any one of claims 17 to 19, wherein said pharmacological agent is selected among chemotherapeutic agents, tyrosine kinase inhibitors, antiproliferative agents, antiemetics, antacids, H2 antagonists, proton pump inhibitors, laxatives, anti-obesity drugs, antidiabetics, vitamins, dietary minerals, antithrombotics, antihemorrhagics, antianginals, antihypertensives, diuretics, vasolidators, beta blockers, calcium channel blockers, rennin-angiotensin system drugs, antihyperlipidemics (statins, fibrates, bile acid sequestrants ) , antipsoriatic, sex hormones, hormonal contraceptives, fertility agents, SERMs, hypothalamic-pituitary hormones, corticosteroids (glucocorticoids, mineralocorticoids) , thyroid hormones/antithyroid agents, antibiotics, antifungals, antimycobacterial, antivirals, vaccines, antiparasitic (antiprotozoals, anthelmintics) , immunomodulators (immunostimulators, immunosuppressants) , anabolic steroids, anti-inflammatories (NSAID) , antirheumatics, corticosteroids, muscle relaxants, bisphosphonate, anesthetics, analgesics, antimigraines, anticonvulsants, mood stabilizers, antiparkinson drug, psycholeptic (anxiolytics, antipsychotics, hypnotics/sedatives) , psychoanaleptic (antidepressants, stimulants/psychostimulants) , decongestants, bronchodilators, Hl antagonists. 21. Process according to any one of claims 17 to 20, wherein said isogenic human cell line is selected among MCFlOA, hTERT-HMEl, HTERT-RPE-I, HCT 116, DLD-I, SW48, NuLi, CuFi, CHON-001, CHON-002, BJ-5ta, hTERT-HMEl (ME16C) , hTERT RPE-I, hTERT-HPNE, NeHepLxHT, T HESCs, RWPE-I, RWPE-2, WPE-stem, WPE-int, WPE1-NA22, WPE1-NB14, WPEl-NBIl, WPE1-NB26, RWPE2-W99, WPMY-I, WPE1-NB26-64 , WPE1-NB26-65, HBE4-E6/E7 [NBE4-E6/E7] , JVM-13, MeT-5A, BBM, BZR, BEAS-2B, MCF 1OA, MCF 1OF, MCF-10-2A, B-3, HBE4-E6/E7-C1 , HK-2, CHON-001, CHON-002, HS-5, PWR-IE, THLE-3, HCE-2 [50. Bl], 46BR. IN, BRISTOL 8, AGLCL, C211, GM1899A, GS-109-V-63, GS-109-V-34, H9, HFFF2, HFLl, HG261, HH-8, HL, Hs 68, Hs 888Lu, HsI. Tes, IM 9, MRC-5 pdl9, MRC-5 pd25, MRC-5 pd30, MRC-5 pd30, MRC-5 SVl TGl, MRC-5 SVl TG2, MRC-5 SV2 , MRC-7, MRC-9, MT-2, PNTlA, PNTlA (SERUM FREE) , PNT2, PNT2 (SERUM FREE) , SVCT, SVCT-MI2, TK6, TK6TGR, TOU (TOU 1-2), WI 26 VA4 , WI 38, WI 38VA13 Subline 2RA, WiDr, WIL2 NS, WIL2.NS.6TG, WILCL, OVCAR-5, OVCAR-4 , OVCAR-3, NCI-H522, NCI-H460, NCI-H322M, NCI-H23, NCI-H226, NCI/ADR-RES, MOLT-4, MDA-N, MDA-MB-435, MDA-MB-231, MCF7, Malme-3M, M14, LOXIMVl, KM12, K-562, IGROVl, HT-29, Hs 578T, HOP-92, HOP-62, HL-60, HCT-15, HCT-116, HCC-2998, EKVX, DU-145, COLO-205, CCRF-CEM, CAKI-I, BT-549, ACHN, A549, A498, 786-0 cell lines. 22. Process according to any one of claims 17 to 21, wherein said cancer allele is selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2, AKT3, KDR, MKK4 , FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (pl6) , BRCAl, BRCA2, PTCHl, VHL, SMAD4 , PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 23. Process according to any one of claims 17 to 22, wherein said at least one mutated cancer allele is selelected among the mutated cancer alleles listed in Table 2a. 24. Cell bank comprising a plurality of isogenic human cell lines, wherein said cell lines comprise at least one mutated cancer allele, wherein said at least one mutated cancer allele is under the control of an endogenous promoter of said cell line, said endogenous promoter being corresponding to the wild-type cancer allele promoter. 25. Cell bank according to claim 24, wherein said cell line carries at least two mutated cancer alleles. 26. Cell bank according to claim 24 or 25, wherein said cell lines are selected among MCFlOA, hTERT-HMEl, HTERT-RPE-I, HCT 116, DLD-I, SW48, NuLi, CuFi, CHON-001, CHON-002, BJ-5ta, hTERT-HMEl (ME16C) , hTERT RPE-I, hTERT-HPNE, NeHepLxHT, T HESCs, RWPE-I, RWPE-2, WPE-stem, WPE-int, WPE1-NA22, WPE1-NB14, WPEl-NBIl, WPE1-NB26, RWPE2-W99, WPMY-1, WPE1-NB26-64, WPE1-NB26-65, HBE4-E6/E7 [NBE4-E6/E7] , JVM-13, MeT-5A, BBM, BZR, BEAS-2B, MCF 10A, MCF 1OF, MCF-10-2A, B-3, HBE4-E6/E7-C1, HK-2, CHON-001, CHON-002, HS-5, PWR-IE, THLE-3, HCE-2 [50. Bl], 46BR. IN, BRISTOL 8, AGLCL, C211, GM1899A, GS-109-V-63, GS-109-V-34, H9, HFFF2, HFLl, HG261, HH-8, HL, Hs 68, Hs 888Lu, HsI. Tes, IM 9, MRC-5 pdl9, MRC-5 pd25, MRC-5 pd30, MRC-5 pd30, MRC-5 SVl TGl, MRC-5 SVl TG2, MRC-5 SV2, MRC-7, MRC-9, MT-2, PNTlA, PNTlA (SERUM FREE), PNT2, PNT2 (SERUM FREE) , SVCT, SVCT-MI2, TK6, TK6TGR, TOU (TOU 1-2) , WI 26 VA4, WI 38, WI 38VA13 Subline 2RA, WiDr, WIL2 NS, WIL2.NS.6TG, WILCL, OVCAR-5, OVCAR-4 , OVCAR-3, NCI-H522, NCI-H460, NCI-H322M, NCI-H23, NCI-H226, NCI/ADR- RES, MOLT-4, MDA-N, MDA-MB-435, MDA-MB-231, MCF7, Malme-3M, M14, LOXIMVl, KM12, K-562, IGROVl, HT-29, Hs 578T, HOP-92, HOP-62, HL-60, HCT-15, HCT-116, HCC-2998, EKVX, DU-145, COLO-205, CCRF-CEM, CAKI-I, BT-549, ACHN, A549, A498, 786-0 cell lines. 27. Cell bank according to any one of claims 24 to 26, wherein said at least one cancer allele is selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, C-KIT, C-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4 , FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (pl6), BRCAl, BRCA2, PTCHl, VHL, SMAD4 , PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 28. Cell bank according to any one of claims 23 to 26, wherein said at least one mutated cancer allele is selelected among the mutated cancer alleles listed in Table 2a. 29. Everolimus for use in the treatment of a patient suffering from a tumor, wherein said tumor carries a mutated PIK3CA cancer allele and is free of a KRAS mutated cancer allele. 30. Indomethacin for use as a medicament in the treatment of a patient suffering from a tumor.
ISOGENIC HUMAN CELL LINES COMPRISING MUTATED CANCER ALLELES AND PROCESS USING THE CELL LINES
BARDELLI, Alberto;DI NICOLANTONIO, Federica;ARENA, Sabrina
2008-01-01
Abstract
Isogenic human cell lines comprising at least one mutated cancer allele under the control of the cell line endogenous promoter, which corresponds to the wild-type cancer allele promoter are disclosed, as well as an in vitro process for determining sensitivity/resistance of a patient suffering from a tumor to a pharmacological agent comprising the following steps: a) identifying at least one mutated cancer allele in a tissue affected by a tumor of said patient; b) providing an isogenic human cell line representative of the tissue, wherein the cell line comprises at least the identified mutated cancer allele, which is under the control of the cell line endogenous promoter corresponding to the wild-type cancer allele promoter; c) putting in contact said cell line with the pharmacological agent; d) determining a variation of proliferation, apoptosis or cytotoxicity of the cell line in presence of the pharmacological agent; wherein the variation of proliferation, apoptosis or cytotoxicity is indicative of the sensitivity/resistance of the patient tumor to the pharmacological agent. Claims 1. Isogenic human cell line, characterized in that said cell line comprises at least one mutated cancer allele, in that said at least one mutated cancer allele is under the control of an endogenous promoter of said cell line, said endogenous promoter being corresponding to the wild-type cancer allele promoter, and in that said cancer allele is selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4, FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (pl6) , BRCAl, BRCA2 , PTCHl, VHL, SMAD4, PERl, MENl, NFl, NF2, ATM, PTPRD. 2. Isogenic human cell line according to claim 1, wherein said cell line carries at least two mutated cancer alleles, wherein said at least two cancer alleles are selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3 , ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4, FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (plβ) , BRCAl, BRCA2 , PTCHl, VHL, SMAD4, PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 3. Isogenic human cell line according to claim 1 or claim 2, wherein said at least one mutated cancer allele is selelected among the mutated cancer alleles listed in Table 2a. 4. Isogenic human cell line according to any one of claims 1 to 3, wherein said human cell line is selected among MCFlOA, hTERT-HMEl, HTERT-RPE-I, HCT 116, DLD-I, SW48, NuLi, CuFi, CHON-001, CHON-002, BJ-5ta, hTERT-HMEl (ME16C) , hTERT RPE-I, hTERT-HPNE, NeHepLxHT, T HESCs, RWPE-I, RWPE-2, WPE-stem, WPE-int, WPE1-NA22, WPE1-NB14, WPEl-NBIl, WPEl-NB26, RWPE2-W99, WPMY-I, WPE1-NB26-64 , WPE1-NB26-65, HBE4-E6/E7 [NBE4-E6/E7] , JVM-13, MeT-5A, BBM, BZR, BEAS-2B, MCF 1OA, MCF 1OF, MCF-10-2A, B-3, HBE4-E6/E7-C1 , HK-2, CHON-001, CHON-002, HS-5, PWR-IE, THLE-3, HCE-2 [50. Bl], 46BR. IN, BRISTOL 8, AGLCL, C211, GM1899A, GS-109-V-63, GS-109-V-34, H9, HFFF2, HFLl, HG261, HH-8, HL, Hs 68, Hs 888Lu, HsI. Tes, IM 9, MRC-5 pdl9, MRC-5 pd25, MRC-5 pd30, MRC-5 pd30, MRC-5 SVl TGl, MRC-5 SVl TG2, MRC-5 SV2 , MRC-7, MRC-9, MT-2, PNTlA, PNTlA (SERUM FREE), PNT2, PNT2 (SERUM FREE) , SVCT, SVCT-MI2, TK6, TK6TGR, TOU (TOU 1-2), WI 26 VA4 , WI 38, WI 38VA13 Subline 2RA, WiDr, WIL2 NS, WIL2.NS.6TG, WILCL, OVCAR-5, OVCAR-4 , OVCAR-3, NCI-H522, NCI-H460, NCI-H322M, NCI-H23, NCI-H226, NCI/ADR-RES, MOLT-4, MDA-N, MDA-MB-435, MDA-MB-231, MCF7, Malme-3M, M14, LOXIMVl, KM12, K-562, IGROVl, HT-29, Hs 578T, HOP-92, HOP-62, HL-60, HCT-15, HCT-116, HCC-2998, EKVX, DU-145, COLO-205, CCRF-CEM, CAKI-I, BT-549, ACHN, A549, A498, 786-0 cell lines. 5. Isogenic human cell line according to any one of the preceding claims, wherein the mutated BRAF cancer allele carries the mutation V600E as shown in SEQ ID No . : 3. 6. Isogenic human cell line according to any one of the preceding claims, wherein the mutated EGFR cancer allele carries the mutation delE746-A750 as shown in SEQ ID No . : 5. 7. Isogenic human cell line according to any one of the preceding claims, wherein the mutated PIK3CA cancer allele carries the mutations E545K and H1047R as shown in SEQ ID No . : 1 and 2, respectively. 8. Isogenic human cell line according to any one of the preceding claims, wherein the mutated CTNNBl cancer allele carries the mutation T41A as shown in SEQ ID No . : 6. 9. Isogenic human cell line according to any one of the preceding claims, wherein the mutated PTEN cancer allele carries the mutation R130* as shown in SEQ ID No . : 7. 10. Isogenic human cell line according to any one of the preceding claims, wherein said cell line carries at least one detectable marker. 11. Isogenic human cell line according to claim 10, wherein said at least one marker is selected among a fluorescent, radioactive, luminescent, phosphorescent marker. 12. Isogenic human cell line according to any one of the preceding claims, wherein said cell line carries at least one knocked-out or inactivated tumor suppressor gene. 13. Isogenic human cell line according to claim 12, wherein said at least one tumor suppressor gene is selected among PTEN, TP53, APC, p21, RbI, BUBl, BRCAl, BRCA2, PTCH, VHL, SMAD4, PERl, TSC2, CDKN2A, DCC, MEN-I, NFl, ATM, PTPRD, LRPlB and NF2. 14. Use of an isogenic human cell line according to any one of claims 1 to 13 for generating xenografts apt to induce tumor growth in a non-human laboratory animal model. 15. Use of an isogenic human cell line according to any one of claims 1 to 13 for producing non-human transgenic laboratory animals susceptible to develop a tumor, said tumor carrying at least one mutated cancer allele, said cancer allele being selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4, FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (plβ) , BRCAl, BRCA2, PTCHl, VHL, SMAD4, PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 16. Use according to claim 15, wherein said non-human transgenic laboratory animals carrying a tumor are used for determining the sensitivity/resistance of said tumor to a pharmacological agent administered to said transgenic animals . 17. In vitro process for determining sensitivity/resistance of a patient suffering from a tumor to a pharmacological agent, characterized in that said process comprises: a) identifying at least one mutated cancer allele in a tissue affected by a tumor of said patient; b) providing an isogenic human cell line representative of said tissue, said cell line comprising at least said mutated cancer allele, wherein said cancer allele is under the control of an endogenous promoter of said cell line, said endogenous promoter being corresponding to the wild-type cancer allele promoter; c) putting in contact said isogenic cell line with said pharmacological agent; d) determining a variation of proliferation, cytotoxicity and/or apoptosis of said isogenic cell line in presence of said pharmacological agent; said variation of proliferation, cytotoxicity and/or apoptosis being indicative of said sensitivity/resistance of said patient to said pharmacological agent. 18. Process according to claim 17, wherein said process further comprises: bl) providing a wild-type isogenic human cell line representative of said tissue, being said wild-type isogenic human cell line free of said mutated cancer allele; cl) putting in contact said wild-type isogenic cell line with said pharmacological agent; dl) determining a variation of proliferation, cytotoxicity and/or apoptosis of said wild-type isogenic cell line in presence of said pharmacological agent. 19. Process according to claim 17 and claim 18, wherein said sensitivity/resistance is evaluated as the relative variation of proliferation, apoptosis and/or cytotoxicity between said isogenic human cell line comprising said at least mutated cancer allele and said wild-type isogenic human cell line. 20. Process according to any one of claims 17 to 19, wherein said pharmacological agent is selected among chemotherapeutic agents, tyrosine kinase inhibitors, antiproliferative agents, antiemetics, antacids, H2 antagonists, proton pump inhibitors, laxatives, anti-obesity drugs, antidiabetics, vitamins, dietary minerals, antithrombotics, antihemorrhagics, antianginals, antihypertensives, diuretics, vasolidators, beta blockers, calcium channel blockers, rennin-angiotensin system drugs, antihyperlipidemics (statins, fibrates, bile acid sequestrants ) , antipsoriatic, sex hormones, hormonal contraceptives, fertility agents, SERMs, hypothalamic-pituitary hormones, corticosteroids (glucocorticoids, mineralocorticoids) , thyroid hormones/antithyroid agents, antibiotics, antifungals, antimycobacterial, antivirals, vaccines, antiparasitic (antiprotozoals, anthelmintics) , immunomodulators (immunostimulators, immunosuppressants) , anabolic steroids, anti-inflammatories (NSAID) , antirheumatics, corticosteroids, muscle relaxants, bisphosphonate, anesthetics, analgesics, antimigraines, anticonvulsants, mood stabilizers, antiparkinson drug, psycholeptic (anxiolytics, antipsychotics, hypnotics/sedatives) , psychoanaleptic (antidepressants, stimulants/psychostimulants) , decongestants, bronchodilators, Hl antagonists. 21. Process according to any one of claims 17 to 20, wherein said isogenic human cell line is selected among MCFlOA, hTERT-HMEl, HTERT-RPE-I, HCT 116, DLD-I, SW48, NuLi, CuFi, CHON-001, CHON-002, BJ-5ta, hTERT-HMEl (ME16C) , hTERT RPE-I, hTERT-HPNE, NeHepLxHT, T HESCs, RWPE-I, RWPE-2, WPE-stem, WPE-int, WPE1-NA22, WPE1-NB14, WPEl-NBIl, WPE1-NB26, RWPE2-W99, WPMY-I, WPE1-NB26-64 , WPE1-NB26-65, HBE4-E6/E7 [NBE4-E6/E7] , JVM-13, MeT-5A, BBM, BZR, BEAS-2B, MCF 1OA, MCF 1OF, MCF-10-2A, B-3, HBE4-E6/E7-C1 , HK-2, CHON-001, CHON-002, HS-5, PWR-IE, THLE-3, HCE-2 [50. Bl], 46BR. IN, BRISTOL 8, AGLCL, C211, GM1899A, GS-109-V-63, GS-109-V-34, H9, HFFF2, HFLl, HG261, HH-8, HL, Hs 68, Hs 888Lu, HsI. Tes, IM 9, MRC-5 pdl9, MRC-5 pd25, MRC-5 pd30, MRC-5 pd30, MRC-5 SVl TGl, MRC-5 SVl TG2, MRC-5 SV2 , MRC-7, MRC-9, MT-2, PNTlA, PNTlA (SERUM FREE) , PNT2, PNT2 (SERUM FREE) , SVCT, SVCT-MI2, TK6, TK6TGR, TOU (TOU 1-2), WI 26 VA4 , WI 38, WI 38VA13 Subline 2RA, WiDr, WIL2 NS, WIL2.NS.6TG, WILCL, OVCAR-5, OVCAR-4 , OVCAR-3, NCI-H522, NCI-H460, NCI-H322M, NCI-H23, NCI-H226, NCI/ADR-RES, MOLT-4, MDA-N, MDA-MB-435, MDA-MB-231, MCF7, Malme-3M, M14, LOXIMVl, KM12, K-562, IGROVl, HT-29, Hs 578T, HOP-92, HOP-62, HL-60, HCT-15, HCT-116, HCC-2998, EKVX, DU-145, COLO-205, CCRF-CEM, CAKI-I, BT-549, ACHN, A549, A498, 786-0 cell lines. 22. Process according to any one of claims 17 to 21, wherein said cancer allele is selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, c-KIT, c-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2, AKT3, KDR, MKK4 , FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (pl6) , BRCAl, BRCA2, PTCHl, VHL, SMAD4 , PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 23. Process according to any one of claims 17 to 22, wherein said at least one mutated cancer allele is selelected among the mutated cancer alleles listed in Table 2a. 24. Cell bank comprising a plurality of isogenic human cell lines, wherein said cell lines comprise at least one mutated cancer allele, wherein said at least one mutated cancer allele is under the control of an endogenous promoter of said cell line, said endogenous promoter being corresponding to the wild-type cancer allele promoter. 25. Cell bank according to claim 24, wherein said cell line carries at least two mutated cancer alleles. 26. Cell bank according to claim 24 or 25, wherein said cell lines are selected among MCFlOA, hTERT-HMEl, HTERT-RPE-I, HCT 116, DLD-I, SW48, NuLi, CuFi, CHON-001, CHON-002, BJ-5ta, hTERT-HMEl (ME16C) , hTERT RPE-I, hTERT-HPNE, NeHepLxHT, T HESCs, RWPE-I, RWPE-2, WPE-stem, WPE-int, WPE1-NA22, WPE1-NB14, WPEl-NBIl, WPE1-NB26, RWPE2-W99, WPMY-1, WPE1-NB26-64, WPE1-NB26-65, HBE4-E6/E7 [NBE4-E6/E7] , JVM-13, MeT-5A, BBM, BZR, BEAS-2B, MCF 10A, MCF 1OF, MCF-10-2A, B-3, HBE4-E6/E7-C1, HK-2, CHON-001, CHON-002, HS-5, PWR-IE, THLE-3, HCE-2 [50. Bl], 46BR. IN, BRISTOL 8, AGLCL, C211, GM1899A, GS-109-V-63, GS-109-V-34, H9, HFFF2, HFLl, HG261, HH-8, HL, Hs 68, Hs 888Lu, HsI. Tes, IM 9, MRC-5 pdl9, MRC-5 pd25, MRC-5 pd30, MRC-5 pd30, MRC-5 SVl TGl, MRC-5 SVl TG2, MRC-5 SV2, MRC-7, MRC-9, MT-2, PNTlA, PNTlA (SERUM FREE), PNT2, PNT2 (SERUM FREE) , SVCT, SVCT-MI2, TK6, TK6TGR, TOU (TOU 1-2) , WI 26 VA4, WI 38, WI 38VA13 Subline 2RA, WiDr, WIL2 NS, WIL2.NS.6TG, WILCL, OVCAR-5, OVCAR-4 , OVCAR-3, NCI-H522, NCI-H460, NCI-H322M, NCI-H23, NCI-H226, NCI/ADR- RES, MOLT-4, MDA-N, MDA-MB-435, MDA-MB-231, MCF7, Malme-3M, M14, LOXIMVl, KM12, K-562, IGROVl, HT-29, Hs 578T, HOP-92, HOP-62, HL-60, HCT-15, HCT-116, HCC-2998, EKVX, DU-145, COLO-205, CCRF-CEM, CAKI-I, BT-549, ACHN, A549, A498, 786-0 cell lines. 27. Cell bank according to any one of claims 24 to 26, wherein said at least one cancer allele is selected among BRAF, EGFR, PIK3CA, PTEN, CTNNBl, C-KIT, C-MET, EPHA3, Erbb2, AKTl, FGFR2, MSH6, ABLl, STATl, STAT4, RET, AKT3, TEK, VAV3, ALK, LYN, NOTCH, IDHl, RORl, FLT3, ALK, SRC, BCL9, RPS6KA2, PDPKl, NTRK3, NTRK2 , AKT3, KDR, MKK4 , FBWX7, MEKl, OBSCN, TECTA, MLL3, NRAS, HRAS, TP53, APC, RbI, CDKN2A (pl6), BRCAl, BRCA2, PTCHl, VHL, SMAD4 , PERl, MENl, NFl, NF2, ATM, PTPRD, KRAS. 28. Cell bank according to any one of claims 23 to 26, wherein said at least one mutated cancer allele is selelected among the mutated cancer alleles listed in Table 2a. 29. Everolimus for use in the treatment of a patient suffering from a tumor, wherein said tumor carries a mutated PIK3CA cancer allele and is free of a KRAS mutated cancer allele. 30. Indomethacin for use as a medicament in the treatment of a patient suffering from a tumor.
| File | Dimensione | Formato | |
|---|---|---|---|
|
Brevetto_WO2010_063300A1.pdf
Accesso riservato
Tipo di file:
POSTPRINT (VERSIONE FINALE DELL’AUTORE)
Dimensione
6.6 MB
Formato
Adobe PDF
|
6.6 MB | Adobe PDF | Visualizza/Apri Richiedi una copia |
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
