Kiran Khush (Stanford Univ CA) "Cell-free DNA in transplant medicine" #NGDx15
5:21pm August 19th 2015 via Hootsuite
Pantel: Q:Functional studies - does extraction bias it? A:Beads can affect CTCs, can't be excluded. Protein exp more robust #NGDx15
5:20pm August 19th 2015 via Hootsuite
Pantel: Q:CTC clusters? A:Some reports of high imptc but low #'s analyzed to-date. They see them, many types, but need to be careful #NGDx15
5:19pm August 19th 2015 via Hootsuite
Pantel: Q:Use of EpCAM vs. newer techniques? A:A sep. talk. They are testing >10 platforms now. #NGDx15
5:18pm August 19th 2015 via Hootsuite
Pantel: Progressive br ca pt who refused therapy - very high CTC (50K/7.5mL) The variable release of DNA as ctDNA needs CTC to compl #NGDx15
5:17pm August 19th 2015 via Hootsuite
Pantel: Cp CTCs and ctDNA - complementary. 2013 review http://t.co/UoKPROQnIl #NGDx15 2011 review http://t.co/bQbhE4tDYf
5:14pm August 19th 2015 via Hootsuite
Pantel: Recent 2015 paper http://t.co/t73yknH1aW established a cell line from CRC CTCs #NGDx15
5:11pm August 19th 2015 via Hootsuite
Pantel: CTCs can initiate metastasis in xenograft assay Nature Biotech '13 http://t.co/KTagY7HS3r #NGDx15
5:10pm August 19th 2015 via Hootsuite
Pantel: Dist of mutations in primary tumor, met and CTC:17 or 20 'private CTC mutations' found. 2013 paper http://t.co/8X99nuMeBN #NGDx15
5:08pm August 19th 2015 via Hootsuite
Pantel: PD1/PDL1 mediated immune blockade - T-cell interaction interrupted. But which benefit? PDL1 on CTC http://t.co/M29RWgJu1i #NGDx15
5:07pm August 19th 2015 via Hootsuite
Pantel: Prostate ca AR-V7 in CTCs from mCRC - this 2014 NEJM paper http://t.co/5YizFDL7tn #NGDx15
5:05pm August 19th 2015 via Hootsuite
Pantel: Doing genotype/phenotype correlation between ER+/ER- CTCs in br ca pts - WGA and sequencing #NGDx15
5:03pm August 19th 2015 via Hootsuite
Pantel: Detection of HER2 oncogene in br ca http://t.co/GZgdY8shXT Now recruiting DETECT-II for anti-HER2 therapy #NGDx15
5:02pm August 19th 2015 via Hootsuite
Pantel: Also referred to this 2012 Cancer Res piece http://t.co/TMye4Cf5ry #NGDx15
5:00pm August 19th 2015 via Hootsuite
Pantel: Began by illustrating different approaches from this 2014 review http://t.co/ZqJMSqpypO #NGDx15
4:59pm August 19th 2015 via Hootsuite
Klaus Pantel (Univ Hamburg Germany) "Circulating cell-free DNA and CTCs as complementary sources of liq biopsy in cancer patients" #NGDx15
4:57pm August 19th 2015 via Hootsuite
Erlander: Q:Assay plex? A:Currently it is single-plex, working on expanding #NGDx15
4:05pm August 19th 2015 via Hootsuite
Erlander: Q:How much plasma used? A:2mL to 4mL #NGDx15
Erlander: Q:After collection, how stable? A:Now up to 14d at RT. They do add a preservative. #NGDx15
4:03pm August 19th 2015 via Hootsuite
Erlander: Q:Monitoring organ transplantation and with damaged kidney fn? A: Not known, guess proportional to GFR #NGDx15
Erlander: Q:Rec'd urine void time? A:Typically morning. Now studying it with later in the day. #NGDx15
4:01pm August 19th 2015 via Hootsuite
Erlander: Also used their urine assay in plasma: assaying KRAS Kaplan-Meier separating low vs high. Prospect of hazard ratio changes #NGDx15
3:58pm August 19th 2015 via Hootsuite
Erlander: Prospective, ongoing study at USC with mCRC, KRAS pos, get chemotherapy 4pts, 4 different responses, in adv of CEA, imging #NGDx15
3:55pm August 19th 2015 via Hootsuite
Erlander: Study with anti-EGFR TKI, monitoring in day 1-3 showing a lot of tumor cells being killed #NGDx15
3:50pm August 19th 2015 via Hootsuite
Erlander: Working with UCSD, onto NSCLC for EGFR T790M resistance mutation. Early data shows earlier detection than imaging. NGDx15
3:48pm August 19th 2015 via Hootsuite
Erlander: 100% found BRAF in urine, only 70% in tissue for systemic histiocytosis Work with MSKCC and MDAnderson #NGDx15
3:45pm August 19th 2015 via Hootsuite
Erlander: Recent prospective study on BRAF V600E with cfDNA in Cancer Disc 2015 http://t.co/2BUGaRlr42 #NGDx15
3:42pm August 19th 2015 via Hootsuite
Erlander: footprint is very short - amplicons are 31bp - 46bp dep on the assay. Data for EGFR Ex19del and KRAS G12D shown #NGDx15
3:39pm August 19th 2015 via Hootsuite
Erlander: Ave urine void is 350mL, their assay requires 100mL. Their assay - suppress wild-type via primer blocker #NGDx15
3:38pm August 19th 2015 via Hootsuite
Erlander: HMW about 2kb-4kb. Use autom. mag-bead-based method for enrichment, that cuts out the HMW material. 10x of cfDNA in urine #NGDx15
3:37pm August 19th 2015 via Hootsuite
Erlander: They started on RainDance, moved onto MiSeq. Extraction - showed distribution of size. Before enrichment: LMW 90, 180bp #NGDx15
3:36pm August 19th 2015 via Hootsuite
Erlander: From blood to kidney to urine: platform comprises of purification, enrichment, detect, quantify. Detect is agnostic #NGDx15
3:35pm August 19th 2015 via Hootsuite
Mark Erlander (Trovagene CA) "Urinary circulating free DNA platform for diagnosis and cancer treatment monitoring" #NGDx15
3:33pm August 19th 2015 via Hootsuite
Siravegna: Q:What is a 'must-have' for clinicians? A:Wide NGS-approach complements targeted. First discover then monitor #NGDx15
3:31pm August 19th 2015 via Hootsuite
Siravegna: Q:How long is HMW DNA in urine? A: LMW is 100bp; HMW is in the kb range #NGDx15
3:28pm August 19th 2015 via Hootsuite
Siravegna: Q:How to decide what to look for in ctDNA? A: 1% mutation is reasonable as mCRC has high burden, and can screen sev genes #NGDx15
3:26pm August 19th 2015 via Hootsuite
Siravegna: Liquid biopsies may move beyond blood to urine - working with Trovagene - low MW and high MW that serves as gDNA ref NGDx15
3:24pm August 19th 2015 via Hootsuite
Siravegna: Pts non-responsive to EGFR blockade, blood-based ID of actionable targets #NGDx15
3:18pm August 19th 2015 via Hootsuite
Siravegna: Recent 2015 Annals of Oncol ref: http://t.co/KrmikJ3ipz looking at early Rx response for mCRC #NGDx15
3:15pm August 19th 2015 via Hootsuite
Siravegna: KRAS, NRAS, BRAF most common mutations screened, 97% plasma-tissue concordance. #NGDx15
3:12pm August 19th 2015 via Hootsuite
Siravegna:Blood samples for ctDNA analysis from diagnosis onward. mCRC - 100 pts, 200 samples (plasma plus matched gDNA). #NGDx15
3:10pm August 19th 2015 via Hootsuite
Siravegna: Describes BEAMing, capturing 1 bead/emulsion, can get 1/10,000 wild-type. ddPCR called 'BEAMing in a box' #NGDx15
3:08pm August 19th 2015 via Hootsuite
Siravegna: Collaborated on Bettegowda et al 2014 STM paper - for many tumor types. NGS is 2% sens; qPCR 1%; BEAMing 0.01%; ddPCR #NGDx15
3:07pm August 19th 2015 via Hootsuite
Siravegna: NRAS, KRAS, EGFR mut's, both intra- and inter-tumor heterogeneity. ctDNA released from secretion, apoptosis or necrosis #NGDx15
3:04pm August 19th 2015 via Hootsuite
Siravegna: In colorectal cancer (CRC) reviewed EGFR signalling block with Cetuximab, resistance mechanisms to EGFR blockade #NGDx15
3:02pm August 19th 2015 via Hootsuite
Giulia Seravegna (Univ Torino Italy) "Blood-based genotyping of colorectal cancer patients" #NGDx15
3:00pm August 19th 2015 via Hootsuite
Rosenfeld: Q:Does sample collection matter? A: Starting studies now. 'Room temp at 24h appears to be okay' 'Streck tubes up to 4d' #NGDx15
2:54pm August 19th 2015 via Hootsuite
Rosenfeld: Q:How is the noise reduced? A: Optimized the up-front prep (8-12 steps) individually to help suppress #NGDx15
2:50pm August 19th 2015 via Hootsuite
Rosenfeld: Q:Input amt? A: Only 200uL blood (about 1ng of DNA) for amplicon. #NGDx15
2:48pm August 19th 2015 via Hootsuite
Rosenfeld:Q:How to handle low frequency mut? A:Below 0.1% - varies per-sample. Says it is in data analysis, limited by amt of sample #NGDx15
