1 / 37

Melanoma Cell Lines in Radiation Therapy Research

Explore the use of melanoma cell lines as a model for experimental radiation therapy and new approaches in biomarkers research. Discover different types of melanoma, stages and survival rates, genetic mutations, and targeted therapy.

jleasure
Télécharger la présentation

Melanoma Cell Lines in Radiation Therapy Research

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. Melanoma cell lines as a model for experimental radiation therapy – new approaches in biomarkers research Cracow, 13th of September, 2018 Prof. Ewa Ł. Stępień Dept. Of Medical Physics, Faculty of Physics Astronomy and Applied Computer Sciences

  2. Types of skin cancer

  3. MELANOCYTES MELANOMA CELL LINE WM-115

  4. Diagnostic alphabet http://drcarrollderm.com/benign-premalignant-malignant-skin-lesions/

  5. Melanoma stages vs. survival rate

  6. Melanoma types • Superficial spreading melanoma -commonly found on the arms, legs, chest and back. The melanoma cells usually grow slowly at first, and spread out across the surface of the skin. • Nodular melanoma - can grow more quickly than other melanomas. It is also more likely to lose its colour when growing, becoming red rather than black. It is more commonly found on the chest, back, head or neck. • Lentigo maligna melanoma - usually found in older people, in areas of skin that have had a lot of sun exposure over many years. It is often found on the face and neck. • Acral lentiginous melanoma - usually found on the palms of the hands, soles of the feet, or under fingernails or toenails. It is more common in people with black or brown skin. It is not thought to be related to sun exposure.

  7. Pie chart demonstrating the percentagedistribution of genes with clinically-relevant and recurrent driver mutations in individuals with melanoma, including non-V600 BRAF alterations (left), interrogated on the Vanderbilt melanoma SNaPshotassay. BRAF fusionsoccur in approximately 4–8% “pan-negative” cases (right).

  8. Frequency of Somatic Gene Mutations in Melanoma (only oncogenic driver mutations)

  9. Supgroups of melanoma show different types of gene mutations INTEMITTENT SUN-EXPOSURE superficial and nodular multiple melanocytic nevi frequent BRAF-mutations rere other mutations CHRONIC SUN-EXPOSURE acral and mucosal melanoma only few melanocytic nevi rarely BRAF-mutations C-Kit mutations

  10. Molecular subsets of melanoma *Frequencies of MEK1 and CTNNB1 mutations according to anatomic location are not known at present

  11. Targeted Therapy for Melanoma Skin Cancer

  12. Targeted Therapy for Melanoma Skin Cancer

  13. Experimental protocol leading to establishment of melanoma cell lines https://histologyblog.com/2014/01/23/histoquarterly-superficial-spreading-malignant-melanoma/ radial/vertical growth phase lymph node metastasis placed in tissue culture flask for in vitro establishment subcutanous implant (xenograft) in mouse Cancer Melanoma cell lines primary CM line WM-115 matastatic CM WM-266-4 primary CM WM-793 Cancer Melanoma cell lines matastatic CM WM-1205Lu

  14. WM-115 - Overview - Cell Line Synopsis

  15. WM-115 - Similarity by Gene Expression Profile

  16. WM-115 - Similarity by Gene mutations Total number of mutated genes: WM-115: 529 WM-266-4: 519 WM-793: 613 T. Hulsen, J. de Vlieg and W. Alkema, BMC Genomics 2008, 9 (1): 488

  17. WM-115 - Similarity by Gene mutations WM-115 vs WM-793 shared mutations  BRAF->ENST00000288602->p.V600E WM-115 vs WM-266-4 shared mutationsBRAF->ENST00000288602->p.V600E WM-115 vs WM-266-mel shared mutationsBRAF->ENST00000288602->p.V600D WM-115 vs UACC-62 shared mutationsBRAF->ENST00000288602->p.V600E

  18. WM-115/WM-266-4 Similarity by somatic gene mutations in cancer ID: BRAFID: DOCK3ID: PXDNLID: DNAH10ID: COL11A1ID: SPTA1ID: PAPPA2ID: KCNT2ID: CACNA1SID: OBSCNID: CNTN5ID: TECTAID: GRIN2BID: KSR2ID: RNF17ID: SIPA1L1ID: UNC13CID: ALPK3ID: PKD1ID: NLRP1ID: DNAH2ID: MYH8ID: DNAH9ID: SCN4AID: MUC16ID: NPHS1ID: MSH6ID: ANKRD36ID: LRP1BID: XIRP2ID: TTNID: PDE1AID: DNAH7ID: PTPRTID: MYO18BID: ERC2ID: CMYA5ID: TNXBID: DSTID: SYNE1ID: SDK1ID: PCLOID: GRM3ID: CSMD1ID: ST18ID: ADCY8ID: PLECID: PRUNE2ID: COL5A1ID: MXRA5ID: CROCCID: CSMD2ID: SCAF1ID: PCDH15 f>8% 25 patients- Mutated Genes 25 profiled samples TCGA T. Hulsen, J. de Vlieg and W. Alkema, BMC Genomics 2008, 9 (1): 488

  19. WM-115/WM-266-4 Similarity by somatic gene mutations in cancer ID: BRAFID: LHCGRID: ADAM28ID: DOCK3ID: PXDNLID: DNAH10ID: GRIK3ID: C8AID: COL11A1ID: SPTA1ID: PAPPA2ID: KCNT2ID: CACNA1SID: OBSCNID: SORCS3ID: TDRD1ID: SLC22A9ID: CNTN5ID: TECTAID: GRIN2BID: SYT10ID: KSR2ID: RNF17ID: MYO16ID: MIA2ID: SIPA1L1ID: FBN1ID: UNC13CID: ALPK3ID: PKD1ID: SRRM2ID: ABCC6ID: DNAH3ID: ITGADID: ZNF423ID: NLRP1ID: DNAH2ID: MYH8ID: MYH3ID: DNAH9ID: NCOR1ID: SCN4AID: BPTFID: DSG3ID: SALL3ID: MUC16ID: ZNF208ID: NPHS1ID: PSG9ID: LRP1BID: XIRP2ID: TTNID: PDE1AID: ZSWIM2ID: DNAH7ID: ABCA12ID: PTPRTID: TRPM2ID: MYO18BID: CACNA1DID: ERC2ID: CCDC158ID: SHROOM3ID: LRBAID: CMYA5ID: TNXBID: DSTID: COL19A1ID: MDN1ID: PTPRKID: SYNE1ID: SDK1ID: PCLOID: GRM3ID: LAMB4ID: CSMD1ID: ST18ID: ADCY8ID: PLECID: DOCK8ID: PRUNE2ID: COL5A1ID: MXRA5ID: CSMD2ID: ANKRD11ID: PCDH15ID: MYCBP2 f>8% 1171 patients- Mutated Genes 1097 profiled samples

  20. Mechanism of BRAF-relatedcancerogenesis

  21. BRAF mutations in melanoma - genome Mutation ID: COSM476 Gene name:   BRAF AA mutation: p.V600E (Substitution - Missense, position 600, V➞E) CDS mutation: c.1799T>A (Substitution, position 1799, T➞A) Nucleotides inserted: n/a Genomic coordinates: GRCh38, 7:140753336..140753336, View: Ensembl contig http://www.ensembl.org/Homo_sapiens/Location/View?g=ENSG00000157764;r=7:140753270-140753400;t=ENST00000288602;db=core;mr=7:140753336-140753337

  22. BRAF mutations in melanoma - trancript Exons: 19, Coding exons: 19, Transcript length: 2,561 bps, Translation length: 806 residues http://www.ensembl.org/Homo_sapiens/Transcript/Summary?db=core;g=ENSG00000157764;mr=7:140753336-140753337;r=7:140753270-140753400;t=ENST00000288602 Serine/threonine-protein kinase B-raf https://cancer.sanger.ac.uk/cosmic3d/protein/BRAF

  23. https://cancer.sanger.ac.uk/cosmic/mutation/overview?id=476 https://cancer.sanger.ac.uk/cosmic3d/protein/BRAF?pdb=4MNE

  24. BRAF Mutations

  25. BRAF mutations in melanoma - pathway https://www.genome.jp/kegg-bin/show_pathway?hsa05218+N00013

  26. Genomics of Drug Sensitivity in Melanoma

  27. Genoskin ex vivo human skin model

  28. Genoskin ex vivo human skin model

  29. In vivospheroids 3D melanoma model 3D melanoma spheroids implanted into a collagen gel matrix  https://www.nature.com/articles/srep19103

  30. In vivospheroids 3D melanoma model AnaSP software (http://sourceforge.net/p/anasp/) is used to automatically compute several morphological parameters (3D reconstructions obtained by using ReViSP, http://sourceforge.net/p/revisp/) https://www.nature.com/articles/srep19103

  31. In vivospheroids 3D melanoma model https://www.nature.com/articles/srep19103

  32. In vitro melanoma model in radiotherapy

  33. Conclusions • WM-115, WM-266-4 and WM0793 melanoma cell lines are representative with regards the oncogenic and clinically-relevant mutations - BRAF • Melanoma cell lines are used for drug testing in 2D systems • 3D systems are proposed for in vitro research to mimic in vivo conditions • 3D systems are proposed to use in radiotherapy testing

  34. Acknowledgements • Sonia Wróbel, PhD student • Ewelina Kubicz, PhD student

  35. McKenzie

More Related