1 / 27

Tumour Modifier Genes

Tumour Modifier Genes. 1. STRATEGY (KISS OF DEATH). 2. SOME RESULTS. 3. WHY DO HUMAN ASSOCIATION STUDIES NOT WORK (WELL)?. RADIATION. CHEMICAL MUTAGEN. TUMOR PROMOTER. SV40T. Ki-ras. Mouse Modifier Program. Skin. Lung. Prostate. Lymphoma. GENETIC APPROACHES TO TUMOR MODIFIERS

ejosefina
Télécharger la présentation

Tumour Modifier Genes

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. Tumour Modifier Genes 1. STRATEGY (KISS OF DEATH) 2. SOME RESULTS 3. WHY DO HUMAN ASSOCIATION STUDIES NOT WORK (WELL)?

  2. RADIATION CHEMICAL MUTAGEN TUMOR PROMOTER SV40T Ki-ras Mouse Modifier Program Skin Lung Prostate Lymphoma

  3. GENETIC APPROACHES TO TUMOR MODIFIERS • INTRASPECIFIC MUSCULUS X MUSCULUS CROSSES • backcross • intercross • recombinant inbred • recombinant congenic • INTERSPECIFIC SPRETUS X MUSCULUS CROSSES • ADVANCED MULTISTRAIN INTERCROSS • 4 OUTBRED POPULATIONS

  4. ADVANTAGES • GENETIC DIVERGENCE - extreme phenotypic differences • DOMINANT RESISTANCE GENES – reduced complexity • 3HIGHLY POLYMORPHIC- mapping is easy DISADVANTAGES • Male F1s are sterile • Can be difficult to breed INTERSPECIFIC SPRETUS-MUSCULUS CROSS

  5. SpGla SPRET/Ei C57/B6 C3H NIH/Ola Gene Expression in Mus spretus and Mus musculus 16 14 12 10 Log2 Control Signal Intensity 8 6 4 2 0

  6. Dominant resistance genes in mus spretus Mus spretus are resistant to tumor development in the SKIN LUNG COLON LIVER LYMPHOID SYSTEM

  7. INTERSPECIFIC BACKCROSS: LINKAGE ANALYSIS AND HAPLOTYPING OUTBRED SPRETUS X B F1 HYBRID: X OR

  8. Human 20q13  Mouse Tumor Susceptibility Loci 17 18 19 X 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 0 Scc6 Par4 Msmr1 Sluc13 Skts9 Sluc7 Rmv1,2,3 Scc8 10 Rmcf Pas4 Rfv1,2 Tgct1 Lts1 Skts5 Ter Lyr2 Rfv3 Pas3 Sluc10 Rrs Sluc14 Gct2 20 Hcf1 Scc5 Skts3 Hcr2 Skts10 Hcs1 Liver Pgct1 Esl1 Hcs6 Scc2 Skts1 Rgv1 Lyr1 Par3 Gct3 30 Msmr2 Gct4 Fv4 Foc1 Foc1 Sluc4 Pas7 Pas6 Ccs1 Skts11 Ril1 Scc4 Pas9 (Pas5) Par2 40 Ots1 Rfv3 Esl1 Hcr1 Pas6 Sluc1 Tlag1 Pas5b Pctr1 Skts6 Scc1 Par1 Hcs5 Papg1 50 Sluc11 Hcs2 Hcs3 Skts7 Sluc12 Sluc9 Sluc6 Lyr3 Sluc3 Fv2 Tlsr2 Scc9 60 Ssic1 Psl1 Skts4 Skts2 Mom1 Pas1 Tli1 Pctr1 Tlsm1 Liver 70 Gct1 Skts12 Sluc8 Pctr2 Pctm Rvil1 Fv1 80 Hcf2 Scc7 Hcs7 Ccs2 Pas8 90 Ril3 Lung loci Skin loci Skts8 Hcs4 Skts13 Scc3 Lung + Skin loci

  9. Microsatellite Haplotyping of outbred spretus The size of the spretus alleles present in the F1BX mice was determined for microsatellite markers spanning the entire distal region of chr 2. Subsequently, haplotypes were constructed for association studies. A single region of 1-2 cM is shared by the haplotypes that are associated with papilloma resistance. The order of all indicated microsatellites was determined and confirmed by recombinant mice.

  10. mouse human

  11. Chromosome 20q13.2 Candidate Locus HEFL ZNF217 STK6 BCAS1 PFD4 Unknown hypothetical protein MCR3 60S Ribosomal Protein Like CYP24 PIC1L Q9NTU7 CSTEF1 Unknown OVC10-2 Unknown 23 kDA 54.5 Mb 56.5 Mb 57.5 Mb 55.5 Mb Amplicons in human tumors

  12. AURORA kinase INVOLVED IN CHROMOSOME SEGREGATION AND ANEUPLOIDY: AMPLIFIED IN HUMAN CANCERS TUMOR STRONG ALLELE WEAK ALLELE “HIGH RISK” “LOW RISK”

  13. Aurora 2 Ile 31 stimulates tumor growth a Number of cells (x104) Days 2 5 0 0 2 0 0 0 1 5 0 0 1 0 0 0 5 0 0 0 1 0 2 0 3 0 Ile31 b Rat1 STK15 Phe-31.1 Rat1 STK15 Phe-31.2 Rat1 STK15 Ile-31.1 Rat1 STK15 Ile-31.2 Phe 31 Rat1 a-Myc c Ile31 Number of cells (x104) Phe 31 Days d rat1 STK15 Phe-31 Ile31 rat1 STK15 Ile-31 Tumor volume cm3 Phe 31 Days Fig. 4

  14. STK15 Phe-31 / hsp70 STK15 Ile-31 / Hsp70 STK15 Phe-31 / UBE2N STK15 Ile-31 / UBE2N Empty vector / UBE2N P53 / T7 Fig. 1

  15. STK15 Phe-31 a STK15 Ile-31 Rec UBE2N beads IP: a-His IB: a-UBE2N Total STK15 10859A 10859A 10859A 07038D 07038D 07038D 10859A lysate 10859A lysate 10859A lysate IP: a-STK15 IP: a-STK15 IP: a-STK15 IB: a-UBE2N IB: a-UBE2N IB: a-UBE2N Total STK15 Total STK15 Total STK15 Total UBE2N Total UBE2N Total UBE2N Aurora 2 Phe-31 preferentially binds UBE2N 293 cells Hom Phe-31 Hom Ile-31 Lymphoblastoid cells

  16. Aurora2 – Phe 31 relocalises UBE2N to the centrosome DAPI a-HA 293 STK15 Phe-31-HA/UBE2N-Myc Merge a-Myc 10mm a-HA DAPI 293 STK15 Ile-31-HA/UBE2N-Myc Merge a-Myc 10mm Fig. 3

  17. GENETIC CHANGES AT TUMOR MODIFIER GENES? NORMAL CELL RESISTANCE SUSCEPTIBILITY TUMOR CELL OR DELETION AMPLIFICATION

  18. 100% 50% % of chromosomal imbalance 90% 80% 0% 70% D5Mit4 D6Mit4 D6Mit9 D9Mit9 D1Mit33 D4Mit12 D5Mit66 D5Mit78 D5Mit81 D6Mit14 D6Mit15 D7Mit69 D7Mit12 D7Mit14 D9Mit76 D12Mit5 D16Mit2 D17Mit7 D1Mit102 D4Mit166 D4Mit160 D6Mit223 D7Mit246 D9Mit179 D9Mit136 D12Mit68 D16Mit88 D12Mit186 D12Mit245 D16Mit102 D17Mit176 D17Mit177 D17Mit139 D17Mit109 Markers 60% 40% 30% 20% 10%

  19.                                                                                                                                               Array CGH Normal DNA-cy5 Co-hybridize to arrayed clones Data analysis Cot-1   Tumor DNA-cy3

  20. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 X K-ras Amplification is an Early Event in the Progression of Lung Tumors in K-rasLA2 Mice

  21. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 X Array CGH on Different Size K-ras Lung Tumors

  22. ALLELE-SPECIFIC EXPRESSION OF MOUSE AURORA2 (NIH/Ola x SPRET/Ei) F1 cell line A5 cell line B9 cDNA cDNA cDNA DNA 0 24 48 DNA 0 24 48 DNA 0 24 48 * CT * * * * * NORMAL TUMOR TUMOR (with amplification)

  23. ALLELE-SPECIFIC AMPLIFICATION OF AURORA KINASE IN COLON CANCER Fig 5. Colon tumor amplification of Stk6 alleles

  24. myc +ve -ve • Genetic interactions • Some genes have +ve or -ve effects apoptosis ras TGFb +ve -ve +ve -ve transformation Growth arrest Growth arrest apoptosis Invasion Metastasis Immunosuppression angiogenesis Why are human association studies so weak?

  25. IDENTIFICATION OF MOUSE AND HUMAN TUMOR SUSCEPTIBILITY GENES HUMAN ASSOCIATION STUDY MOUSE LINKAGE DATA SUSCEPTIBILITY GENE HUMAN: ALLELE-SPECIFIC AMPLIFICATION IN TUMORS MOUSE :ALLELE-SPECIFIC AMPLIFICATION AND EXPRESSION IN TUMORS

  26. ACKNOWLEDGEMENTS Mandy Toland Jian-Hua Mao Jin-wei Yuan Jeff Hsu Di Wu Reyno Delrosario Spiros Linardopoulos- London John de Koning Hiroki Nagase Joe Gray Graeme Hodgson June Chan Jing Ma - Harvard Tyler Jacks – MIT Ponder Lab-Cambridge NCI MOUSE MODELS OF HUMAN CANCER CONSORTIUM

  27. Aurora Kinase and human prostate cancer Aurora/STK6 SNP Cases Controls OR 95%C.I. Age-adjusted OR (95%CI) TT 279 305 1.0 - 1.0 - AT 196 182 1.2 0.9-1.5 1.3 (1.0-1.7) AA 40 28 1.6 0.9-2.6 1.7 (1.0-3.0) (P=0.05)

More Related