循环肿瘤细胞最新进展PPT文件格式下载.ppt

1、CD45（+）and CD66（+）lBasic modes:positive selection mode:anti-EpCAM negative selection mode:anti-CD45 anti-CD66 CTC-iChip systemCombines the strengths of microfluidics for rare cell handing and magnetic-based cell sorting.lMagnetic labeling of cells in blood cells.（pos-vs neg-）lDebulking by separation

2、 of nucleated cells from the enucleated.lAlignment of nucleated cells within a microfluidic channel.lDeflection of magnetically tagged cells into a collection channel.The Special conditions for CTC cultrueBlood samples:patients with metastatic ER（+）Nonadherent culture conditions（critical）Serum-free

3、supplemented EGF and FGFunder hypoxic conditions（4%O2）Establish long-term oligoclonal CTC（sustaimed in vitro 6months）Results of CTC culturel CTC lines can only be generated from patients who were either off therapy or progressing on treatment.and cannot be generated from patients who were responding

4、 to treatment.l For three patients,four additional CTC cell lines were established from blood samples drawn at multiple different time points during therapy.In these cases,CTCs were cultured only when patients were progressing on treatment.Results of CTC culturelCultured CTCs shared cytological feat

5、ures with the matched primary CTCs captured.lConsisitent with standard CTC definitions:CK（+）;CD45（-）;proliferative index30%（Ki67）；initial doubling time:3days to 3weeks.lFive CTC lines retained ER（+）,BRx-07 lost ER expresion,but regained in Xenograft tumor.lCTC-derived tumors shared histological and

6、immunohistochemical features with the matched primary tumor.Gene analysis lRNA sequencing analysis 6 cultured cell lines;13 commonly used breast cancer cell lines;29 unclured single CTCs.lMutation analysis SNaPShot:primary tumor（140 mutations in 25 gene）NGS platform:CTC culture and other mutation（10

7、00 genes）Results of gene analysisESR1（BRx-68）,TP53（BRx-68,61）and KRAS（BRx-42）are universally present in all independent CTC cell lines.PIK3CA,KRAS and FGFR2 are arising mutations.The mutant allele frequences of TP53 and ESR1 are increased upon prolonged culture.Choose drug targets ESR1 and PIK3CA fo

8、r drug sensitivity test.Results of Drug Sensitivity testSome CTC drug sensitivity measurements were concordant with clinical history.Results of Drug Sensitivity（BRx-68）SERMs（tamoxifen）and SERD（raloxifene）were ineffective,alone or combined with inhibitors of PIK3-mTOR（everolimus）.HSP90 inhibitor STA9

9、090 demonstrated cytotoxicity,alone or combinition with SERMs or SERD.A low dose of STA9090 suppressed ER level.?Results of Drug Sensitivity（BRx-07）Cultured CTCs were highly sensitive to PIK3CA and FGFR2 inhibitors.Combinations of PIK3CA and FGFR inhibitors showed cooperatove effects.?iIGFR mitigate

10、 sensitivity to iPI3K;iHSP90 stablize mutant kinases.Results of Drug Sensitivity（BRx-07）Different FGFR mutations have different sensitivity to PIK3CA and FGFR2 inhibitors.ConclusionsStudy patterns of drug susceptibility with CTC,combined with genetic context.Enable monitoring of emerging subclones with alterd mutational and drug sensitivity profiles.DiscussionlSamples:patients with metastatic/PVTT which stage?lMode:negative mode.lDrug targets:mutational gene analysis Liu Zixinwhich stage?biomarkers?AFP?