Diagnosing Emergence of Kinase Inhibitor Resistance on a Microchip

Summary

Principal Investigator: Thomas G Graeber
Abstract: DESCRIPTION (provided by applicant): This project unites investigators who have developed and published a new diagnostic platform with clinical scientists who will guide the implementation of this diagnostic approach in monitoring targeted kinase inhibitor resistance in melanoma. A signaling diagnostic test based on the microfluidic image cytometry (MIC) platform will be optimized for monitoring the clinical evolution of known molecularly defined resistance mechanisms during clinical inhibitor treatment. Namely, the platform will monitor pERK, PDGFR[unreadable], pAKT and the apoptosis reporter cleaved Caspase-3/7 using fine needle aspirate (FNA) biopsies. The clinical utility of these assays will be confirmed by serially monitoring patient's tumors during kinase inhibitor therapy - including multiple measurements made before and during treatment and upon progression. The long-term goal is early clinical detection of resistance mechanisms, and 'in patient-treatment'-based prediction of tumor responsiveness to specific kinase inhibitors based on signaling responses. A key issue in analyzing acquired resistance is the limitation of repeat diagnostic measurement of tumors since it is not always feasible to perform surgical biopsies. This can be overcome by developing a minimally-invasive, fine needle aspirate (FNA)-based approach to characterize progressive tumors. The established and published MIC platform is capable of quantitative, single-cell proteomic analysis of multiple signaling molecules using only 200 to 3,000 cells. In past work, simultaneous measurement of four critical signaling proteins within the PI3K signaling pathway was performed on a panel of 19 human brain tumor biopsies to identify clinically distinct patient subgroups. Together with bioinformatic analysis, the MIC platform provides a robust, enabling, in vitro molecular diagnostic technology for systems pathology analysis and personalized medicine. Activating B-RAFV600E kinase mutations occur in 50% of human melanomas. Early clinical experience with a novel mutant BRAF-selective inhibitor, vemurafenib, have demonstrated an unprecedented 80% anti-tumor response rate among patients with B-RAFV600E-positive melanomas. However, acquired drug resistance frequently develops after initial responses. Recent studies by the UCLA team unveiled that mechanisms of acquired resistance to B-RAF inhibition include i) activating an RTK (PDGFRb)-dependent survival pathway in addition to MAPK, or ii) reactivating the MAPK pathway via N-RAS mutations. This work will develop a MIC-based signaling assay to monitor signaling changes associated with BRAF inhibitor resistance in melanoma. Subsequently, the microfluidic melanoma-signaling assay will be applied to patient fine needle aspirate biopsies pre- and post-treatment with BRAF inhibitors. The particularly aggressive nature of melanoma, and the established clinical and basic science programs of the UCLA melanoma team (T. Graeber, H.-R. Tseng, R. Lo and A. Ribas), make the melanoma microfluidic diagnostic approach uniquely positioned to impact ongoing clinical trials and therapy.
Funding Period: 2012-08-24 - 2014-07-31
more information: NIH RePORT

Top Publications

  1. pmc Inhibition of CSF-1 receptor improves the antitumor efficacy of adoptive cell transfer immunotherapy
    Stephen Mok
    Authors Affiliations Department of Molecular and Medical Pharmacology Jonsson Comprehensive Cancer Center Surgery, Division of Surgical Oncology Institute for Molecular Medicine Departments of Urology and Pediatrics Crump Institute for Molecular Imaging Department of Medicine, Division of Hematology Oncology, University of California, Los Angeles, Los Angeles Plexxikon Inc, Berkeley, California and Roswell Park Cancer Institute, Buffalo, New York
    Cancer Res 74:153-61. 2014
  2. pmc Metastatic castration-resistant prostate cancer reveals intrapatient similarity and interpatient heterogeneity of therapeutic kinase targets
    Justin M Drake
    Department of Microbiology, Immunology, and Molecular Genetics, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, Division of Hematology and Oncology, Department of Medicine, Molecular Biology Institute, Jonsson Comprehensive Cancer Center, Department of Pathology and Laboratory Medicine, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, Institute for Molecular Medicine, California NanoSystems Institute, and Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095
    Proc Natl Acad Sci U S A 110:E4762-9. 2013
  3. pmc Phosphoproteomic analysis of platelets activated by pro-thrombotic oxidized phospholipids and thrombin
    Alejandro Zimman
    Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, United States of America
    PLoS ONE 9:e84488. 2014
  4. ncbi Adenovirus E4ORF1-induced MYC activation promotes host cell anabolic glucose metabolism and virus replication
    Minh Thai
    Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
    Cell Metab 19:694-701. 2014

Research Grants

Detail Information

Publications4

  1. pmc Inhibition of CSF-1 receptor improves the antitumor efficacy of adoptive cell transfer immunotherapy
    Stephen Mok
    Authors Affiliations Department of Molecular and Medical Pharmacology Jonsson Comprehensive Cancer Center Surgery, Division of Surgical Oncology Institute for Molecular Medicine Departments of Urology and Pediatrics Crump Institute for Molecular Imaging Department of Medicine, Division of Hematology Oncology, University of California, Los Angeles, Los Angeles Plexxikon Inc, Berkeley, California and Roswell Park Cancer Institute, Buffalo, New York
    Cancer Res 74:153-61. 2014
    ..In conclusion, CSF-1R blockade with PLX3397 improved the efficacy of ACT immunotherapy by inhibiting the intratumoral accumulation of immunosuppressive macrophages...
  2. pmc Metastatic castration-resistant prostate cancer reveals intrapatient similarity and interpatient heterogeneity of therapeutic kinase targets
    Justin M Drake
    Department of Microbiology, Immunology, and Molecular Genetics, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, Division of Hematology and Oncology, Department of Medicine, Molecular Biology Institute, Jonsson Comprehensive Cancer Center, Department of Pathology and Laboratory Medicine, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, Institute for Molecular Medicine, California NanoSystems Institute, and Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095
    Proc Natl Acad Sci U S A 110:E4762-9. 2013
    ..Phosphoproteomic analyses and identification of kinase activation states in metastatic castration-resistant prostate cancer patients have allowed for the prioritization of kinases for further clinical evaluation. ..
  3. pmc Phosphoproteomic analysis of platelets activated by pro-thrombotic oxidized phospholipids and thrombin
    Alejandro Zimman
    Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, United States of America
    PLoS ONE 9:e84488. 2014
    ..Our results provide multiple insights into the mechanism of platelet activation and specifically in platelet regulation by oxPCCD36. ..
  4. ncbi Adenovirus E4ORF1-induced MYC activation promotes host cell anabolic glucose metabolism and virus replication
    Minh Thai
    Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
    Cell Metab 19:694-701. 2014
    ..Our findings show how a viral protein exploits host cell machinery to reprogram cellular metabolism and promote optimal progeny virion generation...

Research Grants30

  1. Targeting Pten - An Upstream, Downstream and Offstream Approach
    ZE apos EV A RONAI; Fiscal Year: 2013
    ..abstract_text> ..
  2. CELLULAR AND MOLECULAR MECHANISMS OF GASTROINTESTINAL CANCERS
    Lopa Mishra; Fiscal Year: 2013
    ..Significantly, this program promises to yield new therapies targeted at these difficult-to-treat lethal cancers at the bench, and then to translate the results rapidly into clinical care, at the bedside. ..
  3. Signaling in Inflammation, Stress, and Tumorigenesis
    GEORGE ROBERT STARK; Fiscal Year: 2013
    ..abstract_text> ..
  4. Molecular and Functional Analysis of Single Circulating Melanoma Cells
    Hsian Rong Tseng; Fiscal Year: 2013
    ....
  5. CSHL CANCER CENTER SUPPORT GRANT
    BRUCE W STILLMAN; Fiscal Year: 2013
    ....
  6. Core Center for Musculoskeletal Disorders
    Louis J Soslowsky; Fiscal Year: 2013
    ..abstract_text> ..
  7. Model-based predictions of responses RTK Pathway therapies
    Joe W Gray; Fiscal Year: 2013
    ..abstract_text> ..
  8. Molecular approaches to overcome intrinsic drug resistance in BRAF and NRAS-mutan
    Jessie Villanueva; Fiscal Year: 2013
    ..Collectively, this knowledge will be critical to develop new strategies that eliminate nearly all melanoma cells and inform the design of future clinical trials. ..
  9. DF/HCC Kidney Cancer SPORE
    David McDermott; Fiscal Year: 2013
    ..The overall goal of the DF/HCC Kidney Cancer SPORE is the translation of biological and technological advances into clinically meaningful advances for patients with kidney cancer. ..