GENETAG TECHNOLOGY, INC — Department of Health and Human Services SBIR Phase I: 102

Description

Super sensitive detection of EGFR mutants driving lung cancer recurrence Confidential Principal InvestigatorShaferDavid APhD PROJECT SUMMARYCancer is the leading cause of death worldwideand lung cancer is the leading cause of cancer related deathSeveral point mutations or deletions in the EGFR geneLR exondel codonsexonare common in non small cell lung carcinomaNSCLCand exhibit high response rates to first generation tyrosine kinase inhibitorsTKIsDrug resistance commonly occurs following expansion of a clone sharboring the EGFR TM mutationwhich is often present at low levels before targeted therapy is initiatedbut can also arise during treatmentSeveral third generation TKIs have been developed that irreversibly inhibit TMHoweverthird generation TKIs fail after the emergence of subclones harboring one of andgtadditional EGFR mutationssuch as CSLQor LVTreatment outcomes can be influenced by whether TM is present in cissame alleleor transdifferent alleleswith respect to variants conferring resistance to thirdgeneration TKIsRecentlya fourth generation allosteric inhibitor of TM and CS was reportedCollectivelythese findings indicate that effective NSCLC therapy depends upon the presence and cis trans status of first third generation TKI resistance mutationsboth before treatment and during remissionThe overall objective of this Phase I SBIR project is to develop a PCR basedliquid biopsy assay for detecting circulating EGFR mutations in NSCLC patients that confer drug resistanceand thus lead to recurrenceGeneTAG Technology develops molecular diagnostic assays for cancer and infectious diseasesOur primary systemthe internal DNA Detection SwitchiDDSprobe systemcomprises two interacting componentsa fluor labeled probeand a quencher labeled antiprobe that is nearly complementary to the probeIn the absence of the intended targetthe paired probes and antiprobes bind togetherquenching fluorescence and preventing off target detectionThis unique probe system shows superior single base discrimination over a wider annealing temperature rangeCthan common methodsRecentlywe merged our iDDS probe technology with our Wild TerminatorWTxmethods that enable detection of rare mutantsfrequencyby blocking amplification of the wild type sequenceIn pilot studiesthe combined method showed greater sensitivity in detecting circulating EGFR variants from lung cancer patients than the FDA approved cobasEGFR Mutation TestvThis enhanced qPCR sensitivity meets or exceeds the sensitivity of specialized platforms without a high instrumentation costOur higher sensitivity and specificity will enable development of a platform independentliquid biopsy assayOur Specific Aims areto develop XNA enhanced WTx iDDS probe assays for detecting EGFR drug resistance mutationsandto study drug resistance EGFR mutations in plasma from NSCLC patients in remissionSuccess in Phase I will justify expanded Phase II assay validation studies in preparation for filing for FDA approvalMore sensitive detection against an expanded set of EGFR drug resistance mutations associated with recurrence will guide physicians in selecting first line therapy Super sensitive detection of EGFR mutants driving lung cancer recurrence Confidential Principal InvestigatorShaferDavid APhD PROJECT NARRATIVE Cancer is the leading cause of death worldwideand small mutations in the EGFR gene are known to drive many lung cancers or to be a source of drug resistanceWe have invented highly precise probes for detecting small DNA mutations and are developing companion methods that greatly enhance probe sensitivityWe have also developed specific probes and selective amplification methods for a key set of EGFR mutations that relate to lung cancerWe expect to further develop and apply these methods to detect a series of drug resistance mutations that are associated with recurrence using blood samples from non small cancer lung carcinoma patientsThese advances will be particularly valuable for cancer diagnostics since they offer the potential for liquid biopsy assayswherein solid tumorssuch as in lung or brain tissuemight be assessed non invasively and more readily in small blood samples