JP6683745B2 - 膵管腺癌のサブタイピングのための新規バイオマーカー - Google Patents
膵管腺癌のサブタイピングのための新規バイオマーカー Download PDFInfo
- Publication number
- JP6683745B2 JP6683745B2 JP2018019195A JP2018019195A JP6683745B2 JP 6683745 B2 JP6683745 B2 JP 6683745B2 JP 2018019195 A JP2018019195 A JP 2018019195A JP 2018019195 A JP2018019195 A JP 2018019195A JP 6683745 B2 JP6683745 B2 JP 6683745B2
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- subtype
- pdac
- hnf
- vitro method
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 201000008129 pancreatic ductal adenocarcinoma Diseases 0.000 title claims description 114
- 239000000101 novel biomarker Substances 0.000 title description 5
- 108010020382 Hepatocyte Nuclear Factor 1-alpha Proteins 0.000 claims description 62
- 102100022057 Hepatocyte nuclear factor 1-alpha Human genes 0.000 claims description 62
- 108090000623 proteins and genes Proteins 0.000 claims description 61
- 230000014509 gene expression Effects 0.000 claims description 57
- 238000000034 method Methods 0.000 claims description 40
- 102100028340 Keratin, type II cuticular Hb1 Human genes 0.000 claims description 36
- 101710115634 Keratin, type II cuticular Hb1 Proteins 0.000 claims description 36
- 206010028980 Neoplasm Diseases 0.000 claims description 30
- 238000000338 in vitro Methods 0.000 claims description 30
- 239000003550 marker Substances 0.000 claims description 29
- 238000001514 detection method Methods 0.000 claims description 27
- 102100021374 Hepatocyte nuclear factor 3-gamma Human genes 0.000 claims description 22
- 101000818741 Homo sapiens Hepatocyte nuclear factor 3-gamma Proteins 0.000 claims description 22
- 102000004169 proteins and genes Human genes 0.000 claims description 20
- 108010061414 Hepatocyte Nuclear Factor 1-beta Proteins 0.000 claims description 18
- 102100022123 Hepatocyte nuclear factor 1-beta Human genes 0.000 claims description 18
- 102100024152 Cadherin-17 Human genes 0.000 claims description 13
- 101710196881 Cadherin-17 Proteins 0.000 claims description 13
- 239000012634 fragment Substances 0.000 claims description 13
- 102100029284 Hepatocyte nuclear factor 3-beta Human genes 0.000 claims description 12
- 101001062347 Homo sapiens Hepatocyte nuclear factor 3-beta Proteins 0.000 claims description 12
- 108091023040 Transcription factor Proteins 0.000 claims description 12
- 102000040945 Transcription factor Human genes 0.000 claims description 12
- 102100022047 Hepatocyte nuclear factor 4-gamma Human genes 0.000 claims description 11
- 102100029087 Hepatocyte nuclear factor 6 Human genes 0.000 claims description 11
- 101001045749 Homo sapiens Hepatocyte nuclear factor 4-gamma Proteins 0.000 claims description 11
- 101000988619 Homo sapiens Hepatocyte nuclear factor 6 Proteins 0.000 claims description 11
- 239000000090 biomarker Substances 0.000 claims description 10
- 230000027455 binding Effects 0.000 claims description 9
- 238000011895 specific detection Methods 0.000 claims description 9
- 108020004999 messenger RNA Proteins 0.000 claims description 7
- 230000001105 regulatory effect Effects 0.000 claims description 7
- 238000011529 RT qPCR Methods 0.000 claims description 6
- 108091034117 Oligonucleotide Proteins 0.000 claims description 5
- 210000004923 pancreatic tissue Anatomy 0.000 claims description 5
- 238000011534 incubation Methods 0.000 claims description 4
- 230000002285 radioactive effect Effects 0.000 claims description 4
- 241000124008 Mammalia Species 0.000 claims description 3
- 238000011532 immunohistochemical staining Methods 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 238000005481 NMR spectroscopy Methods 0.000 claims description 2
- 239000007850 fluorescent dye Substances 0.000 claims description 2
- 101000634537 Homo sapiens Neuronal PAS domain-containing protein 2 Proteins 0.000 claims 3
- 102100029045 Neuronal PAS domain-containing protein 2 Human genes 0.000 claims 3
- -1 C110RF9 Proteins 0.000 claims 2
- 101150072844 APOM gene Proteins 0.000 claims 1
- 102100027211 Albumin Human genes 0.000 claims 1
- 102100039951 Annexin A13 Human genes 0.000 claims 1
- 102100037324 Apolipoprotein M Human genes 0.000 claims 1
- 108010036280 Aquaporin 4 Proteins 0.000 claims 1
- 102000012002 Aquaporin 4 Human genes 0.000 claims 1
- 102100035683 Axin-2 Human genes 0.000 claims 1
- 102100038495 Bile acid receptor Human genes 0.000 claims 1
- 102100028736 Claudin-10 Human genes 0.000 claims 1
- 102100024484 Codanin-1 Human genes 0.000 claims 1
- 102100035595 Cohesin subunit SA-2 Human genes 0.000 claims 1
- 102100033635 Collectrin Human genes 0.000 claims 1
- 108700040183 Complement C1 Inhibitor Proteins 0.000 claims 1
- 102100027309 Cyclic AMP-responsive element-binding protein 5 Human genes 0.000 claims 1
- 102100038418 Cytoplasmic FMR1-interacting protein 2 Human genes 0.000 claims 1
- 102100027100 Echinoderm microtubule-associated protein-like 4 Human genes 0.000 claims 1
- 102100027300 Extracellular serine/threonine protein kinase FAM20C Human genes 0.000 claims 1
- 102100021654 Extracellular sulfatase Sulf-2 Human genes 0.000 claims 1
- 102100026339 F-box-like/WD repeat-containing protein TBL1X Human genes 0.000 claims 1
- 102100027844 Fibroblast growth factor receptor 4 Human genes 0.000 claims 1
- 108700031843 GRB7 Adaptor Proteins 0.000 claims 1
- 101150052409 GRB7 gene Proteins 0.000 claims 1
- 108010001496 Galectin 2 Proteins 0.000 claims 1
- 102100039558 Galectin-3 Human genes 0.000 claims 1
- 102100037260 Gap junction beta-1 protein Human genes 0.000 claims 1
- 102100039684 Glucose-6-phosphate exchanger SLC37A4 Human genes 0.000 claims 1
- 102100033107 Growth factor receptor-bound protein 7 Human genes 0.000 claims 1
- 102100022664 Guanylate cyclase activator 2B Human genes 0.000 claims 1
- 102100033961 Guanylin Human genes 0.000 claims 1
- 102100031496 Heparan sulfate N-sulfotransferase 2 Human genes 0.000 claims 1
- 101000693913 Homo sapiens Albumin Proteins 0.000 claims 1
- 101000959674 Homo sapiens Annexin A13 Proteins 0.000 claims 1
- 101000874569 Homo sapiens Axin-2 Proteins 0.000 claims 1
- 101000603876 Homo sapiens Bile acid receptor Proteins 0.000 claims 1
- 101000776612 Homo sapiens Cilia- and flagella-associated protein 73 Proteins 0.000 claims 1
- 101000766993 Homo sapiens Claudin-10 Proteins 0.000 claims 1
- 101000980888 Homo sapiens Codanin-1 Proteins 0.000 claims 1
- 101000642968 Homo sapiens Cohesin subunit SA-2 Proteins 0.000 claims 1
- 101000945075 Homo sapiens Collectrin Proteins 0.000 claims 1
- 101000726193 Homo sapiens Cyclic AMP-responsive element-binding protein 5 Proteins 0.000 claims 1
- 101000956870 Homo sapiens Cytoplasmic FMR1-interacting protein 2 Proteins 0.000 claims 1
- 101001057929 Homo sapiens Echinoderm microtubule-associated protein-like 4 Proteins 0.000 claims 1
- 101000866302 Homo sapiens Excitatory amino acid transporter 3 Proteins 0.000 claims 1
- 101000937709 Homo sapiens Extracellular serine/threonine protein kinase FAM20C Proteins 0.000 claims 1
- 101000820626 Homo sapiens Extracellular sulfatase Sulf-2 Proteins 0.000 claims 1
- 101000835691 Homo sapiens F-box-like/WD repeat-containing protein TBL1X Proteins 0.000 claims 1
- 101001065295 Homo sapiens Fas-binding factor 1 Proteins 0.000 claims 1
- 101000917134 Homo sapiens Fibroblast growth factor receptor 4 Proteins 0.000 claims 1
- 101000954104 Homo sapiens Gap junction beta-1 protein Proteins 0.000 claims 1
- 101000899814 Homo sapiens Guanylate cyclase activator 2B Proteins 0.000 claims 1
- 101001068469 Homo sapiens Guanylin Proteins 0.000 claims 1
- 101000588595 Homo sapiens Heparan sulfate N-sulfotransferase 2 Proteins 0.000 claims 1
- 101001035951 Homo sapiens Hyaluronan-binding protein 2 Proteins 0.000 claims 1
- 101000840630 Homo sapiens Hydroxyproline dehydrogenase Proteins 0.000 claims 1
- 101001081567 Homo sapiens Insulin-like growth factor-binding protein 1 Proteins 0.000 claims 1
- 101000975003 Homo sapiens Kallistatin Proteins 0.000 claims 1
- 101001008946 Homo sapiens Kinesin-like protein KIF12 Proteins 0.000 claims 1
- 101000941884 Homo sapiens Leucine-rich repeat flightless-interacting protein 2 Proteins 0.000 claims 1
- 101001039191 Homo sapiens Leucine-rich repeat-containing protein 19 Proteins 0.000 claims 1
- 101001003687 Homo sapiens Lipoma-preferred partner Proteins 0.000 claims 1
- 101001012669 Homo sapiens Melanoma inhibitory activity protein 2 Proteins 0.000 claims 1
- 101000968663 Homo sapiens MutS protein homolog 5 Proteins 0.000 claims 1
- 101000636823 Homo sapiens Neogenin Proteins 0.000 claims 1
- 101001109685 Homo sapiens Nuclear receptor subfamily 5 group A member 2 Proteins 0.000 claims 1
- 101000596119 Homo sapiens Plastin-3 Proteins 0.000 claims 1
- 101001126417 Homo sapiens Platelet-derived growth factor receptor alpha Proteins 0.000 claims 1
- 101000577585 Homo sapiens Proline dehydrogenase 1, mitochondrial Proteins 0.000 claims 1
- 101001099887 Homo sapiens Rab-3A-interacting protein Proteins 0.000 claims 1
- 101000665882 Homo sapiens Retinol-binding protein 4 Proteins 0.000 claims 1
- 101000581155 Homo sapiens Rho GTPase-activating protein 12 Proteins 0.000 claims 1
- 101000692933 Homo sapiens Ribonuclease 4 Proteins 0.000 claims 1
- 101000650588 Homo sapiens Roundabout homolog 3 Proteins 0.000 claims 1
- 101000654696 Homo sapiens Semaphorin-4G Proteins 0.000 claims 1
- 101000872580 Homo sapiens Serine protease hepsin Proteins 0.000 claims 1
- 101000700735 Homo sapiens Serine/arginine-rich splicing factor 7 Proteins 0.000 claims 1
- 101000864806 Homo sapiens Serine/threonine-protein kinase Sgk2 Proteins 0.000 claims 1
- 101000864831 Homo sapiens Serine/threonine-protein kinase Sgk3 Proteins 0.000 claims 1
- 101000923531 Homo sapiens Sodium/potassium-transporting ATPase subunit gamma Proteins 0.000 claims 1
- 101000596543 Homo sapiens THAP domain-containing protein 11 Proteins 0.000 claims 1
- 101000653005 Homo sapiens Thromboxane-A synthase Proteins 0.000 claims 1
- 101000837626 Homo sapiens Thyroid hormone receptor alpha Proteins 0.000 claims 1
- 101000835726 Homo sapiens Transcription elongation factor A protein 3 Proteins 0.000 claims 1
- 101000819088 Homo sapiens Transcription factor GATA-6 Proteins 0.000 claims 1
- 101001137337 Homo sapiens Transcriptional activator protein Pur-alpha Proteins 0.000 claims 1
- 101000801209 Homo sapiens Transducin-like enhancer protein 4 Proteins 0.000 claims 1
- 101000658581 Homo sapiens Transmembrane 4 L6 family member 4 Proteins 0.000 claims 1
- 101000800288 Homo sapiens Tubulointerstitial nephritis antigen Proteins 0.000 claims 1
- 101000820294 Homo sapiens Tyrosine-protein kinase Yes Proteins 0.000 claims 1
- 101000777220 Homo sapiens Ubiquitin carboxyl-terminal hydrolase 3 Proteins 0.000 claims 1
- 101000772888 Homo sapiens Ubiquitin-protein ligase E3A Proteins 0.000 claims 1
- 101000934581 Homo sapiens Valacyclovir hydrolase Proteins 0.000 claims 1
- 102100039238 Hyaluronan-binding protein 2 Human genes 0.000 claims 1
- 102100027636 Insulin-like growth factor-binding protein 1 Human genes 0.000 claims 1
- 102100023012 Kallistatin Human genes 0.000 claims 1
- 102000001626 Kazal Pancreatic Trypsin Inhibitor Human genes 0.000 claims 1
- 108010093811 Kazal Pancreatic Trypsin Inhibitor Proteins 0.000 claims 1
- 102100027616 Kinesin-like protein KIF12 Human genes 0.000 claims 1
- 102100032694 Leucine-rich repeat flightless-interacting protein 2 Human genes 0.000 claims 1
- 102100040687 Leucine-rich repeat-containing protein 19 Human genes 0.000 claims 1
- 102100026358 Lipoma-preferred partner Human genes 0.000 claims 1
- 102100029778 Melanoma inhibitory activity protein 2 Human genes 0.000 claims 1
- 102000047724 Member 2 Solute Carrier Family 12 Human genes 0.000 claims 1
- 102100032280 Metal cation symporter ZIP14 Human genes 0.000 claims 1
- 102100021156 MutS protein homolog 5 Human genes 0.000 claims 1
- 108010071382 NF-E2-Related Factor 2 Proteins 0.000 claims 1
- 102100031900 Neogenin Human genes 0.000 claims 1
- 102100027341 Neutral and basic amino acid transport protein rBAT Human genes 0.000 claims 1
- 102100031701 Nuclear factor erythroid 2-related factor 2 Human genes 0.000 claims 1
- 102100022669 Nuclear receptor subfamily 5 group A member 2 Human genes 0.000 claims 1
- 102100024894 PR domain zinc finger protein 1 Human genes 0.000 claims 1
- 108090000882 Peptidyl-Dipeptidase A Proteins 0.000 claims 1
- 102000004270 Peptidyl-Dipeptidase A Human genes 0.000 claims 1
- 102100027637 Plasma protease C1 inhibitor Human genes 0.000 claims 1
- 102100035220 Plastin-3 Human genes 0.000 claims 1
- 102100030485 Platelet-derived growth factor receptor alpha Human genes 0.000 claims 1
- 108010009975 Positive Regulatory Domain I-Binding Factor 1 Proteins 0.000 claims 1
- 102100028772 Proline dehydrogenase 1, mitochondrial Human genes 0.000 claims 1
- 102100038475 Rab-3A-interacting protein Human genes 0.000 claims 1
- 102100038246 Retinol-binding protein 4 Human genes 0.000 claims 1
- 102100027663 Rho GTPase-activating protein 12 Human genes 0.000 claims 1
- 102100026411 Ribonuclease 4 Human genes 0.000 claims 1
- 102100027488 Roundabout homolog 3 Human genes 0.000 claims 1
- 101150097162 SERPING1 gene Proteins 0.000 claims 1
- 108091006620 SLC12A2 Proteins 0.000 claims 1
- 102000012979 SLC1A1 Human genes 0.000 claims 1
- 108091006924 SLC37A4 Proteins 0.000 claims 1
- 108091006944 SLC39A14 Proteins 0.000 claims 1
- 108091006946 SLC39A5 Proteins 0.000 claims 1
- 108091006311 SLC3A1 Proteins 0.000 claims 1
- 108091006262 SLC4A4 Proteins 0.000 claims 1
- 108091006277 SLC5A1 Proteins 0.000 claims 1
- 108091006239 SLC7A9 Proteins 0.000 claims 1
- 101150063267 STAT5B gene Proteins 0.000 claims 1
- 102100032781 Semaphorin-4G Human genes 0.000 claims 1
- 102100034801 Serine protease hepsin Human genes 0.000 claims 1
- 102100029287 Serine/arginine-rich splicing factor 7 Human genes 0.000 claims 1
- 102100030069 Serine/threonine-protein kinase Sgk2 Human genes 0.000 claims 1
- 102100024474 Signal transducer and activator of transcription 5B Human genes 0.000 claims 1
- 102000006633 Sodium-Bicarbonate Symporters Human genes 0.000 claims 1
- 102000058090 Sodium-Glucose Transporter 1 Human genes 0.000 claims 1
- 102100034351 Sodium/potassium-transporting ATPase subunit gamma Human genes 0.000 claims 1
- 102100035063 THAP domain-containing protein 11 Human genes 0.000 claims 1
- 102100030973 Thromboxane-A synthase Human genes 0.000 claims 1
- 102100028702 Thyroid hormone receptor alpha Human genes 0.000 claims 1
- 102100026427 Transcription elongation factor A protein 3 Human genes 0.000 claims 1
- 102100021382 Transcription factor GATA-6 Human genes 0.000 claims 1
- 102100035715 Transcriptional activator protein Pur-alpha Human genes 0.000 claims 1
- 102100033763 Transducin-like enhancer protein 4 Human genes 0.000 claims 1
- 102100034897 Transmembrane 4 L6 family member 4 Human genes 0.000 claims 1
- 102100033470 Tubulointerstitial nephritis antigen Human genes 0.000 claims 1
- 102100021788 Tyrosine-protein kinase Yes Human genes 0.000 claims 1
- 102100040198 UDP-glucuronosyltransferase 1-6 Human genes 0.000 claims 1
- 102100029152 UDP-glucuronosyltransferase 1A1 Human genes 0.000 claims 1
- 101710205316 UDP-glucuronosyltransferase 1A1 Proteins 0.000 claims 1
- 101710008381 UGT1A6 Proteins 0.000 claims 1
- 102100031287 Ubiquitin carboxyl-terminal hydrolase 3 Human genes 0.000 claims 1
- 102100030434 Ubiquitin-protein ligase E3A Human genes 0.000 claims 1
- 102100025139 Valacyclovir hydrolase Human genes 0.000 claims 1
- 102100023142 Zinc transporter ZIP5 Human genes 0.000 claims 1
- 102100021298 b(0,+)-type amino acid transporter 1 Human genes 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 42
- 239000000523 sample Substances 0.000 description 23
- 229940122924 Src inhibitor Drugs 0.000 description 19
- 102100035071 Vimentin Human genes 0.000 description 16
- 108010065472 Vimentin Proteins 0.000 description 16
- 210000005048 vimentin Anatomy 0.000 description 16
- 230000035945 sensitivity Effects 0.000 description 12
- 238000013517 stratification Methods 0.000 description 12
- 210000004881 tumor cell Anatomy 0.000 description 12
- 239000003153 chemical reaction reagent Substances 0.000 description 10
- 239000013610 patient sample Substances 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 9
- 238000010195 expression analysis Methods 0.000 description 8
- 239000003814 drug Substances 0.000 description 7
- 238000004393 prognosis Methods 0.000 description 7
- 239000000427 antigen Substances 0.000 description 6
- 108091007433 antigens Proteins 0.000 description 6
- 102000036639 antigens Human genes 0.000 description 6
- 238000013459 approach Methods 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000002493 microarray Methods 0.000 description 5
- 239000002751 oligonucleotide probe Substances 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 210000000496 pancreas Anatomy 0.000 description 4
- 208000008443 pancreatic carcinoma Diseases 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 208000005443 Circulating Neoplastic Cells Diseases 0.000 description 3
- 108010086512 Hepatocyte Nuclear Factor 1 Proteins 0.000 description 3
- 102000006754 Hepatocyte Nuclear Factor 1 Human genes 0.000 description 3
- 108060003951 Immunoglobulin Proteins 0.000 description 3
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 3
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 3
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 3
- 102000001332 SRC Human genes 0.000 description 3
- 108060006706 SRC Proteins 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 102000018358 immunoglobulin Human genes 0.000 description 3
- 238000003364 immunohistochemistry Methods 0.000 description 3
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 3
- 201000002528 pancreatic cancer Diseases 0.000 description 3
- 239000012188 paraffin wax Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- WZRJTRPJURQBRM-UHFFFAOYSA-N 4-amino-n-(5-methyl-1,2-oxazol-3-yl)benzenesulfonamide;5-[(3,4,5-trimethoxyphenyl)methyl]pyrimidine-2,4-diamine Chemical compound O1C(C)=CC(NS(=O)(=O)C=2C=CC(N)=CC=2)=N1.COC1=C(OC)C(OC)=CC(CC=2C(=NC(N)=NC=2)N)=C1 WZRJTRPJURQBRM-UHFFFAOYSA-N 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 2
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 238000001574 biopsy Methods 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 230000009871 nonspecific binding Effects 0.000 description 2
- 210000004940 nucleus Anatomy 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 210000002536 stromal cell Anatomy 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 1
- 102100022900 Actin, cytoplasmic 1 Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 102000000905 Cadherin Human genes 0.000 description 1
- 108050007957 Cadherin Proteins 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 238000009007 Diagnostic Kit Methods 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102000003964 Histone deacetylase Human genes 0.000 description 1
- 108090000353 Histone deacetylase Proteins 0.000 description 1
- 101001045751 Homo sapiens Hepatocyte nuclear factor 1-alpha Proteins 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- 239000005551 L01XE03 - Erlotinib Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 101710192597 Protein map Proteins 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 102000052575 Proto-Oncogene Human genes 0.000 description 1
- 108700020978 Proto-Oncogene Proteins 0.000 description 1
- 102000007696 Proto-Oncogene Proteins c-yes Human genes 0.000 description 1
- 108010021833 Proto-Oncogene Proteins c-yes Proteins 0.000 description 1
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 102000007962 Type II Keratins Human genes 0.000 description 1
- 108010089374 Type II Keratins Proteins 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000013523 data management Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 210000003890 endocrine cell Anatomy 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- AAKJLRGGTJKAMG-UHFFFAOYSA-N erlotinib Chemical compound C=12C=C(OCCOC)C(OCCOC)=CC2=NC=NC=1NC1=CC=CC(C#C)=C1 AAKJLRGGTJKAMG-UHFFFAOYSA-N 0.000 description 1
- 229960001433 erlotinib Drugs 0.000 description 1
- 210000002907 exocrine cell Anatomy 0.000 description 1
- 208000021045 exocrine pancreatic carcinoma Diseases 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 1
- 229960005277 gemcitabine Drugs 0.000 description 1
- 238000010199 gene set enrichment analysis Methods 0.000 description 1
- 230000000762 glandular Effects 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 230000007773 growth pattern Effects 0.000 description 1
- 210000003780 hair follicle Anatomy 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000000370 laser capture micro-dissection Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 238000002966 oligonucleotide array Methods 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 238000002616 plasmapheresis Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 230000020347 spindle assembly Effects 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 210000005166 vasculature Anatomy 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/112—Disease subtyping, staging or classification
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/4742—Keratin; Cytokeratin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/56—Staging of a disease; Further complications associated with the disease
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Hematology (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Biomedical Technology (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Description
本発明は、膵管腺癌 (pancreatic ductal adenocarcinoma)(PDAC)のサブタイプの同定及び層別化のための新規手法に関する。本発明は特に、新規のPDACサブタイプ特異的マーカーに関し、そして前記マーカーを検出するための試薬を含む診断キットに関する。
個別化腫瘍学は、将来癌患者が治療される方法に革命をもたらす可能性を秘めている。実体の異なる癌は、治療応答と臨床的転帰をしばしば決定するシグナル伝達の特異的活性化を含む分子的差異に基づいて、サブクラスに分割することができる。乳癌、肺癌および結腸癌を含む種々の癌について、そのようなサブタイプの同定やコホートへの患者の層別化は、既にサブタイプ特異的な方法で患者を治療するために臨床診療に応用されている。
したがって、本発明の課題は、PDACの、具体的には古典的サブタイプ、間葉系様サブタイプ及び外分泌様サブタイプの新規バイオマーカーを提供することである。それは、PDAC標品の特異的PDACサブタイプへの信頼性ある且つ正確な帰属を可能とする。かかる新規PDACサブタイプ特異的バイオマーカーは、迅速且つ信頼性のある患者の層別化に関する大きな必要性を満たし得る。その結果、サブタイプ特異的な薬剤脆弱性を利用する新規PDAC治療手法の導入及び予後評価を大きく改善する。
驚くべきことに、PDACのサブタイプに特異的な新規バイオマーカーが同定され得ることが明らかとなった。さらに、驚くべきことに、本発明の新規PDACサブタイプ特異的バイオマーカーは、PDAC標品の特異的PDACサブタイプへの信頼性ある且つ正確な帰属を可能とすることが明らかとなった。本発明の新規サブタイプ特異的バイオマーカーは、患者の予後評価を大きく改善するために、及び、サブタイプ特異的な薬剤脆弱性の利用によるPDAC治療手法を大きく改善するために有用であり得る。迅速且つ信頼性のあるPDAC患者層別化は、本発明の新規バイオマーカーを利用することによって初めて可能となった。
本発明は、本明細書中の以下の発明の詳細な説明及び実施例を参照することによってより容易に理解され得る。
一次異種移植片として、原初の腫瘍サブタイプに関連する遺伝子発現パターンを保持しているためサブタイプ特異的バイオマーカーの同定を目的として古典的PDACサブタイプと間葉系様PDACサブタイプと外分泌様PDACサブタイプの分子的特徴づけが可能な商品名付きの安定なPDAC細胞系と、細胞系由来の腫瘍を使用し、サブタイプ特異的遺伝子発現分析を実施した。この遺伝子発現分析においてサブタイプ特異的発現の大きな違い(5倍超)を示す遺伝子が、サブタイプ特異的バイオマーカータンパク質のためのスクリーンに含まれていた。臨床設定では免疫組織化学的マーカーが最も有用であるため、タンパク質地図データベースを使用してこの最初の候補リストを改良した。それに加え、GSEAモチーフモジュールを使用して転写因子のサブタイプ特異的発現の検索も実施した。これにより、外分泌様サブタイプにおいてだけ、転写因子HNF−1のための結合部位を含む遺伝子が豊富であることが明らかになった。したがって外分泌様サブタイプが最終候補リストに含まれた。細胞系由来異種移植片のパネル全体の切片について、あらゆる候補の発現を免疫組織化学法で試験した。腫瘍試料は10%ホルマリンの中で一晩固定し、パラフィンに包埋した。免疫組織化学法のため、スライドを脱パラフィン化し、再水和した。pH6(Dako標的回収溶液、Dako、Glostrup)の蒸気釜中で15分間沸騰させて抗原を回収し、30分間放冷し、蒸留水で洗浄した。Linarisアビジン/ビオチンブロッキングキット(Vector Labs、Burlingame)を製造業者の説明書に従って使用して、非特異的結合をブロックした。スライドを一次抗体と30分間インキュベートし、PBS-T(0.5%トゥイーン-20を含むPBS)で洗浄し、Dako REAL Detection Systemを使用して適切な二次抗体と20分間インキュベートし、PBS-Tで洗浄した。内因性ペルオキシダーゼをブロックし、ストレプトアビジンHRPとインキュベーション(室温で20分間)した後、スライドをAEC(Dako)で発色させ、ヘマトキシリンで対染色した。個々の抗体がPDAC試料の特定のサブセットを染色し、したがって3種類のPDACサブタイプの識別と分類に役立つことがわかった場合には、候補マーカータンパク質だけを選択した。HNF−1Aに対する抗体は、外分泌様サブタイプの腫瘍細胞の核を特異的に染色した。それに対してケラチン81の発現は、間葉系様サブタイプの腫瘍においてだけ検出できた。古典的タイプでは、患者試料を分析するときに予測性があることが後に示される可能性のある特異的マーカーを同定することができなかった。したがって古典的サブタイプのPDACは、ケラチン81とHNF−1Aの発現不在を特徴とすることができる。
miRNAeasyキット(Qiagen、Hilden)を使用してPDAC細胞系又は腫瘍異種移植片から全RNAを単離した。Illmina BeadChip Technology(HumanHT-12)を利用して遺伝子発現分析を実施した。各試料について得られた規格化された遺伝子リストを検出されたシグナルに従って分類した。シグナルが最も強い遺伝子を最上位にし、残りは降順で分類した。ランク付けしたこの遺伝子リストをGSEA-アルゴリズム(Subramanian et al.上記引用)のための入力として用いた。ランク付けした各リストを表1に記載した感度予測因子(SRC-SP)と別々に比較し、FDRを計算した。0.200というFDRカットオフ値が、試料をSrc阻害剤感受性であるかSrc阻害剤抵抗性であるかに正確に分類するのに最適であると判断した。FDRが0.200未満になった試料は感受性であることが予測されるのに対し、FDRが0.200よりも大きいと抵抗性であることが予測される。
我々は、外分泌様サブタイプで過剰発現している転写因子HNF−1の標的遺伝子を同定した。これら遺伝子は、外分泌様サブタイプのためのさらなるマーカーとして用いることができよう。
ホルマリン固定パラフィン包埋組織試料(3μm〜5μmの切片)を脱パラフィン化し、そして再水和した。pH6(Dako標的回収溶液、Dako, Glostrup)の蒸気釜中で15分間沸騰させて抗原を回収し、30分間放冷し、蒸留水で洗浄した。Linaris アビジン/ビオチンブロッキングキット (Vector Labs, Burlingame) を製造業者の説明書に従って使用して、非特異結合をブロックした。スライドを一次抗体と30分間インキュベートし、PBS−T(0.5%ツイーン20を有するPBS)で洗浄し、Dako REAL Detection Systemを使用して適切な二次抗体と20分間インキュベートし、PBS−Tで洗浄した。内因性ペルオキシダーゼをブロックし、ストレプトアビジンHRPとインキュベーション(室温で20分間)した後、スライドをAEC(Dako)で発色させ、ヘマトキシリンで対染色した。染色は、2つの別々の切片に、又は1つの切片上の2つの別々の領域に、それぞれHNF−1Aおよびケラチン81について行った。
ケラチン81陽性/HNF−1A陰性:間葉系様サブタイプ
ケラチン81陰性/HNF−1A陽性:外分泌様サブタイプ
ケラチン81陰性/HNF−1A陰性:古典的サブタイプ
ケラチン81陽性/HNF−1A陽性:未定
サブタイプ特異的な治療手法と組合せた患者の層別化はますます重要になってきており、すでにいくつかのタイプの癌で治療の有効性を改善することが証明されている。しかしこれまで、PDAC患者を臨床的に意味のある群に層別化する試みは、まちまちの結果を与えている(Stathis, A., and Moore, M. J. (2010). Advanced pancreatic carcinoma: current treatment and future challenges. Nat Rev Clin Oncol 7, 163-172)。これは、単一のマーカーによる手法を複雑にする、少なくとも3つのサブタイプの存在に一部起因しているかも知れない。2つのマーカーのセットでは、PACOモデルですべての3つのPDACサブタイプを明確に特定することができる。258人のPDAC患者のコホートへのこれらのマーカーの適用は、非重複染色を確認しており、患者群に適用された時の我々の2つのマーカーセットの頑強性を示唆している。これは、我々が患者の大きなコホートを3つの群[HNF1A陽性(外分泌様)、ケラチン81(KRT81)陽性(QM−PDA)、及び2重陰性(古典的)]に層別化することを可能にした。我々は、3群間で全体的生存率の有意差を見いだした。KRT81陽性患者は最悪の予後(平均OS=16.5ヶ月)を有し、HNF−1A陽性患者は最良の結果(平均OS=43.5ヶ月)を有し、2重陰性患者は中間の結果(平均OS=26.3ヶ月)を有していた。従って我々のマーカーは、PDAC患者の臨床的に意味のある層別化を初めて可能にし、容易に日常的な診断に取り込むことができ、PDAC患者の層別化が、現在の及び新規治療法の指針となる可能性を提供することができる。
Claims (17)
- 腫瘍試料における膵管腺癌(PDAC)細胞のサブタイプ特異的検出のためのインビトロの方法であって、サブタイプ特異的マーカーの発現を、タンパク質又はかかるタンパク質をコードするオリゴヌクレオチドのいずれかとして検出するステップによって特徴づけられ、
ここで、ケラチン81は、間葉系様サブタイプのPDAC細胞のためのマーカーであり、
HNF-1Aは、外分泌様サブタイプのPDAC細胞のためのマーカーであり、そして
古典的サブタイプのPDAC細胞は、ケラチン81及びHNF-1Aの欠如によって特徴づけられる、
前記方法。 - 古典的サブタイプのPDACが、(i)HNF-1A、HNF-1B、FOXA2(HNF3B)、FOXA3(HNF3G)、HNF4G、及びONECUT1(HNF6)から選択される転写因子、(ii)HNF-1Aによって制御される1又は2以上の標的遺伝子、及び(iii) カドヘリン17(CDH17)から選択される1又は2以上の追加のバイオマーカーの特異的発現の欠如を決定することによって同定される、請求項1に記載のインビトロの方法。
- HNF-1Aによって制御される1又は2以上の標的遺伝子が、SLC5A1、FGFR4、ANXA13、C110RF9、TM4SF4、KIF12、RNASE4、UGT1A1、TCEA3、CDH17、SLC4A4、NR5A2、LGALS2、NR1H4、UGT1A6、GATA6、NPAS2、NFE2L2、FAM20C、TBXAS1、IGFBP1、CDAN1、GUCA2B、SLC3A1、FOXA2、STAT5B、MSH5、APOM、FXYD2、HPN、SEMA4G、ROBO3、SLC39A14、SLC7A9、NFIX、PDGFRA、RAB3IP、GC、ANPEP、CRB3、MSX2、MMP11、TLE4、SFRS7、SGK2、GUCA2A、SERPINA4、ALB、GJB1、SLC37A4、SPINK1、SLC12A2、TBL1X、SLC1A1、RBP4、ARHGAP12、NDST2、EML4、NEO1、THRA、PRDM1、LPP、PLS3、YES1、AFM、C14ORF138、BACE2、UBE3A、LRRC19、AQP4、LRRFIP2、BPHL、C5、CYFIP2、SLC39A5、AXIN2、TMEM27、SULF2、MIA2、STAG2、TINAG、USP3、HABP2、CREB5、CLDN10、PURA、PRODH2、GRB7、AGT、TTR、SERPING1、MLL、及びTHAP11から選択される、1又は2以上のHNF-1A標的遺伝子である、請求項2に記載の方法。
- 前記検出が、前記サブタイプ特異的マーカーの1つのエピトープへの検出抗体の結合に基づく、請求項1又は2に記載のインビトロの方法。
- 以下のステップ:
(a)前記サブタイプ特異的マーカーの1つに特異的な検出抗体が前記エピトープと結合できる条件下にて、前記試料を前記検出抗体と接触させ;
(b)過剰の検出抗体分子を除去し;そして
(c)前記試料中で結合した検出抗体を検出すること
を含む、請求項4に記載のインビトロの方法。 - 前記検出抗体は、モノクローナル抗体;ポリクローナル抗体;組み換えによって産生されたキメラ抗体又はヒト化抗体を含む、組み換えによって産生された抗体;完全合成抗体;又は、前記抗体の1つの機能的断片である、請求項4又は5に記載のインビトロの方法。
- 検出抗体の結合は、第二抗体とのインキュベーションによって検出される、請求項4〜6のいずれか1項に記載のインビトロの方法。
- 前記第二抗体は標識化された第二抗体である、請求項7に記載のインビトロの方法。
- 前記第二抗体は、マーカー分子で標識化される、請求項7又は8に記載のインビトロの方法。
- 前記第二抗体は、蛍光染料、生物学的に活性な酵素標識、放射活性標識、核磁気共鳴に活性な標識、発光性標識又は発色団による標識を用いて標識化される、請求項9に記載のインビトロの方法。
- 前記放射活性標識が、32P、125I、3H、14C、188Rh、131I、99mTc及び111Inからなる群から選択される、請求項10に記載のインビトロの方法。
- 抗体の結合が、免疫組織化学染色によって検出される、請求項11に記載のインビトロの方法。
- 前記検出が、前記サブタイプ特異的マーカーの1つをコードするmRNAを測定することに基づく、請求項1〜3のいずれか1項に記載のインビトロの方法。
- 前記検出が、定量的リアルタイムポリメラーゼ連鎖反応(qRT-PCR)によって前記サブタイプ特異的マーカーの1つをコードするmRNAを測定することに基づく、請求項13に記載のインビトロの方法。
- 前記試料は膵臓組織を含む、請求項1〜14のいずれか1項に記載のインビトロの方法。
- 前記試料は、哺乳類から得られる、請求項15に記載のインビトロの方法。
- 前記哺乳類がヒトである、請求項16に記載のインビトロの方法。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP12007128 | 2012-10-12 | ||
EP12007128.7 | 2012-10-12 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015536020A Division JP2015534648A (ja) | 2012-10-12 | 2013-10-14 | 膵管腺癌のサブタイピングのための新規バイオマーカー |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2018105876A JP2018105876A (ja) | 2018-07-05 |
JP6683745B2 true JP6683745B2 (ja) | 2020-04-22 |
Family
ID=47071057
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015536020A Pending JP2015534648A (ja) | 2012-10-12 | 2013-10-14 | 膵管腺癌のサブタイピングのための新規バイオマーカー |
JP2018019195A Active JP6683745B2 (ja) | 2012-10-12 | 2018-02-06 | 膵管腺癌のサブタイピングのための新規バイオマーカー |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015536020A Pending JP2015534648A (ja) | 2012-10-12 | 2013-10-14 | 膵管腺癌のサブタイピングのための新規バイオマーカー |
Country Status (6)
Country | Link |
---|---|
US (2) | US20150260721A1 (ja) |
EP (1) | EP2906953B1 (ja) |
JP (2) | JP2015534648A (ja) |
CA (1) | CA2887920C (ja) |
HK (1) | HK1213640A1 (ja) |
WO (1) | WO2014056626A1 (ja) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016061252A1 (en) * | 2014-10-14 | 2016-04-21 | The University Of North Carolina At Chapel Hill | Methods and compositions for prognostic and/or diagnostic subtyping of pancreatic cancer |
EP3931318A4 (en) * | 2019-04-01 | 2022-12-07 | The University of North Carolina at Chapel Hill | TUMOR PURITY INDEPENDENT SUBTYPING (PURIST), SAMPLE TYPE INDEPENDENT SINGLE SAMPLE PLATFORM AND CLASSIFIER FOR TREATMENT DECISION MAKING IN PANCREATIC CANCER |
EP3730941A1 (en) * | 2019-04-23 | 2020-10-28 | Institut Jean Paoli & Irène Calmettes | Method for determining a reference tumor aggressiveness molecular gradient for a pancreatic ductal adenocarcinoma |
EP4200446A1 (en) * | 2020-08-22 | 2023-06-28 | The Broad Institute, Inc. | Pancreatic ductal adenocarcinoma signatures and uses thereof |
WO2024039196A1 (ko) * | 2022-08-18 | 2024-02-22 | 주식회사 메드팩토 | 암의 진단, 전이가능성 또는 예후 예측용 바이오 마커 및 이의 용도 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE60113287D1 (de) * | 2000-06-14 | 2005-10-13 | Univ Texas | Systeme und verfahren zur zellteilbevölkerungsanalyse |
US20030180747A1 (en) * | 2001-10-11 | 2003-09-25 | Hruban Ralph H. | Pancreatic cancer diagnosis and therapies |
EP1380644A1 (en) * | 2002-07-08 | 2004-01-14 | Kylix B.V. | The use of specified TCF target genes to identify drugs for the treatment of cancer, in particular colorectal cancer, in which TCF/beta-catenin/WNT signalling plays a central role |
US8178505B2 (en) * | 2005-01-05 | 2012-05-15 | Sloan-Kettering Institute For Cancer Research | Method of predicting and reducing risk of metastasis of breast cancer to lung |
US20080026481A1 (en) * | 2006-06-16 | 2008-01-31 | Michael Mitas | Molecular based two-marker assays that predict outcome of adenocarcinoma patients |
JP5240739B2 (ja) * | 2007-04-13 | 2013-07-17 | オーエスアイ・フアーマシユーテイカルズ・エル・エル・シー | キナーゼ阻害剤に対する抗癌応答を予測する生物学的マーカー |
EP2361315B1 (en) * | 2008-11-27 | 2014-03-05 | INSERM - Institut National de la Santé et de la Recherche Médicale | Cxcl4l1 as a biomarker of pancreatic cancer |
CN102414562B (zh) * | 2009-03-24 | 2019-05-07 | 生物概念股份有限公司 | 细胞捕获和分析的装置和方法 |
EP2419177A1 (en) * | 2009-04-17 | 2012-02-22 | OSI Pharmaceuticals, LLC | Biological markers predictive of anti-cancer response to epidermal growth factor receptor kinase inhibitors |
EP2723898A4 (en) * | 2011-06-22 | 2015-09-30 | Oncocyte Corp | METHOD AND COMPOSITIONS FOR THE TREATMENT AND DIAGNOSIS OF BUBBLE CANCER |
-
2013
- 2013-10-14 EP EP13780067.8A patent/EP2906953B1/en active Active
- 2013-10-14 US US14/434,036 patent/US20150260721A1/en not_active Abandoned
- 2013-10-14 WO PCT/EP2013/003085 patent/WO2014056626A1/en active Application Filing
- 2013-10-14 CA CA2887920A patent/CA2887920C/en active Active
- 2013-10-14 JP JP2015536020A patent/JP2015534648A/ja active Pending
-
2016
- 2016-02-16 HK HK16101665.4A patent/HK1213640A1/zh unknown
-
2018
- 2018-02-06 JP JP2018019195A patent/JP6683745B2/ja active Active
-
2019
- 2019-05-24 US US16/422,778 patent/US20190317097A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
JP2015534648A (ja) | 2015-12-03 |
JP2018105876A (ja) | 2018-07-05 |
EP2906953A1 (en) | 2015-08-19 |
CA2887920C (en) | 2021-03-23 |
WO2014056626A1 (en) | 2014-04-17 |
HK1213640A1 (zh) | 2016-07-08 |
US20150260721A1 (en) | 2015-09-17 |
CA2887920A1 (en) | 2014-04-17 |
US20190317097A1 (en) | 2019-10-17 |
EP2906953B1 (en) | 2017-12-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6683745B2 (ja) | 膵管腺癌のサブタイピングのための新規バイオマーカー | |
JP2002504354A (ja) | 癌の治療法 | |
KR101690249B1 (ko) | 암의 진단 방법 및 암 환자의 전반적 생존 및 무질병 생존의 측정 방법 | |
JP5893037B2 (ja) | Brafv600eに特異的に結合する抗体を使用する癌の診断のための手段及び方法 | |
Popov et al. | Prognostic value of EGF receptor and tumor cell proliferation in bladder cancer: therapeutic implications | |
US20100081666A1 (en) | Src activation for determining cancer prognosis and as a target for cancer therapy | |
AU2010274581B2 (en) | A method of diagnosing cancer | |
JP2014519818A (ja) | 前立腺癌用の予測バイオマーカ | |
WO2011000573A1 (en) | Diagnostic method for predicting the risk of cancer recurrence based on histone macroh2a isoforms | |
US20200141938A1 (en) | Methods and Diagnostics for Cancer Detection and Treatment Monitoring | |
US20210148911A1 (en) | Methods and Diagnostics for Cancer Detection and Treatment Monitoring | |
US7588904B2 (en) | Perinucleolar compartment as a cancer marker | |
JP7114112B2 (ja) | 肺癌の予後バイオマーカー | |
US20080274461A1 (en) | Perinucleolar Compartment Markers for Cancer | |
KR102271401B1 (ko) | Romo1를 측정하는 제제를 포함하는 방사선 저항성 또는 감수성 예측용 조성물 및 이의 용도 | |
EP2269073B1 (en) | A method of assessing cancer status in a breast cancer patient | |
US8241862B2 (en) | Perinucleolar compartment as a cancer marker | |
KR20150012090A (ko) | 항-안지오포이에틴 2 항체에 대한 임상 마커 및 이의 이용 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20180308 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20180308 |
|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20181221 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20190108 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20190404 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20190425 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20190903 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20191024 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20200225 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20200326 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6683745 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |