Continuous emulsion digital PCR (cdPCR) for low-cost, rapid, near point-of-care genotyping of non-small cell lung cancer

PROJECT SUMMARY Genotyping of cancer has become the standard of carewhere specific mutations in cancer driving genes have been shown to indicate sensitivity or resistance to targeted treatmentsSampling of cell free DNAcfDNAin blood plasma could provide a non invasive means of performing this genotypingbut current methods are too complexslowdaysand expensivehundreds of dollarsto make use of this information in a manner that would have a major clinical impa ctTo widely impact cancer carea minimally invasive monitoring assay must be inexpensive enough$that it can be ordered repeatedlyand must be fast enoughandlthrsthat it can seamlessly integrate into the current pace of clinical decision makingThis Phase I STTR proposal seeks to evaluate the feasibility of DropWorkscontinuous emulsion digital PCRcdPCRtechnology for rapidlow costnon invasive genotyping of cancer through measurement of cell free DNA in blood plasmaThe high level of automation in this technology greatly reduces the complexity of the workflowallowing it to be placed in typical hospital laboratories and obviating the requirement for specialty laboratory staffIts simplifiedcontinuous nature has the potential to greatly reduce turnaround timeminutesand cost$This tool could provide clinicians with low costhigh frequency information on genotype status and trends for a wide range of mutations known to be indicative of therapeutic responseimproving outcomes through clearer selection and adaptation of treatmentThe project objective will be accomplished in partnership with Dana Farber Cancer Institutewhich has developed a set of validated cfDNA genotyping assays for non small cell lung cancer using digital droplet PCRIn this projectassays for exdeletionLRand TM of EGFR will be the focusas these have specific near term relevance to treatmentThe project will determine whether the existing cdPCR technology can be automatedreducing clinical expertise requirements and costand adapted to DFCI s specific cancer assaysAimand whether the specificity and sensitivity of the assays meet those for the validated digital PCR approach in constructed samplesAimand a small human pilot trialAimA Phaseproposal would extend this work by transforming the instrument into a robust clinical toolconfirming sensitivity and specificity for NSCLC in a wider human trialdesigning a full clinical trialand expanding the assay set to other mutations and cancers PROJECT NARRATIVE This project aims to evaluate the feasibility of the continuous emulsion digital PCRcdPCRtechnology for rapidandlthrlow cost$non invasive genotyping of cancer through the sampling and analysis of cell free DNA in blood plasmaGenotypic information obtained from cell free DNA could provide indication of susceptibility or emerging resistance to targeted therapeuticsbut current methods for analysis are too complexslowand costly to be fast and frequent enough to integrate into clinical decision making and impact outcomesIf successfulthis technology could provide a platform for making serialnon invasive genotyping a routine part of care for a wide range of cancerspositively impacting outcomes for a vast number of patients