Pharmacodynamic Response of the MET/HGF Receptor to Small-Molecule Tyrosine Kinase Inhibitors Examined with Validated, Fit-for-Clinic Immunoassays

Srivastava, Apurva K.; Hollingshead, Melinda G.; Weiner, Jennifer; Navas, Tony; Khin, Sonny; Ji, Jiuping Jay; Zhang, Yiping; Borgel, Suzanne; Pfister, Thomas D.; Kinders, Robert J.; Bottaro, Donald P.; Linehan, W. Marston; Tomaszewski, Joseph E.; Doroshow, James H.; Parchment, Ralph E.

doi:10.1158/1078-0432.ccr-15-2323

Cited by 31 publications

(43 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition to difficulties in obtaining fresh tumor tissue to establish that MET diagnostics reflect the most recent status of cancer patients, two technical problems have hampered translation of pMET measurements in clinical studies: (I) sampling errors due to the limited stability of phospho-signals in biopsies; and (II) availability of specific antibodies and reagents that can be moved beyond the research setting into clinical laboratories. We recently described novel reagents and assays that the NCI has developed, standardized, and validated to facilitate the transition from the research lab to the clinic (13). In this review, we further elaborate on the underlying principles that guided development of our novel MET assays and the unique advantages they offer in providing tangible, quantifiable evidence of MET dysregulation and its blockade by MET inhibitors.…”

Section: Introductionmentioning

confidence: 99%

“…These studies could not identify MET peptides, likely because the enrichment process was insufficient to capture extremely low-abundance proteins. We studied the impact of both cold and warm ischemia time on SNU5 tumors and demonstrated that biopsies must be cryopreserved within three minutes of collection to preserve pMET signals (13). Even a modest delay of 15 minutes under warm/cold ischemia can result in >60% loss in pMET signal.…”

Section: Introductionmentioning

confidence: 99%

“…Details of biopsy collection and processing required for valid measurement of pMET and other PD biomarkers are described in SOP341205 at http://dctd.cancer.gov/ ResearchResources/ResearchResources-biomarkers.htm. Fortunately, once cryopreserved, MET is fairly stable at assay operating temperatures in tissue lysates containing broad-spectrum phosphatase and protease inhibitors for the duration of analysis (13).…”

Section: Introductionmentioning

confidence: 99%

“…In experimental xenograft tumors, this variability dictates that drug-induced changes in pMET levels must exceed 45% in magnitude to be considered distinguishable from the variability due to normal biological heterogeneity and sampling error (13). There are many underlying reasons for this variability.…”

Section: Introductionmentioning

confidence: 99%

“…We employed a total MET immunoassay that not only measures the full-length MET protein levels, but can also be used to normalize the measurement of MET phosphorylation. This allows for reporting of pMET signals as the ratio of pMET to total MET protein, potentially overcoming errors in tumor sampling (13).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Effective implementation of novel MET pharmacodynamic assays in translational studies

et al. 2017

Self Cite

View full text Add to dashboard Cite

MET tyrosine kinase (TK) dysregulation is significantly implicated in many types of cancer. Despite over 20 years of drug development to target MET in cancers, a pure anti-MET therapeutic has not yet received market approval. The failure of two recently concluded phase III trials point to a major weakness in biomarker strategies to identify patients who will benefit most from MET therapies. The capability to interrogate oncogenic mutations in MET via circulating tumor DNA (ctDNA) provides an important advancement in identification and stratification of patients for MET therapy. However, a wide range in type and frequency of these mutations suggest there is a need to carefully link these mutations to MET dysregulation, at least in proof-of-concept studies. In this review, we elaborate how we can utilize recently developed and validated pharmacodynamic biomarkers of MET not only to show target engagement, but more importantly to quantitatively measure MET dysregulation in tumor tissues. The MET assay endpoints provide evidence of both canonical and non-canonical MET signaling, can be used as "effect markers" to define biologically effective doses (BEDs) for molecularly targeted drugs, confirm mechanism-of-action in testing combination of drugs, and establish whether a diagnostic test is reporting MET dysregulation. We have established standard operating procedures for tumor biopsy collections to control preanalytical variables that have produced valid results in proof-of-concept studies. The reagents and procedures are made available to the research community for potential implementation on multiple platforms such as ELISA, quantitative immunofluorescence assay (qIFA), and immuno-MRM assays.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Effective implementation of novel MET pharmacodynamic assays in translational studies

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

Met Signaling in Carcinogenesis

Silva

Roy

Kato

et al. 2018

Predictive Biomarkers in Oncology

View full text Add to dashboard Cite

Combination therapy with pazopanib and tivantinib modulates VEGF and c-MET levels in refractory advanced solid tumors

et al. 2021

Self Cite

View full text Add to dashboard Cite

SummaryThe vascular endothelial growth factor (VEGF)/VEGFR and hepatocyte growth factor (HGF)/c-MET signaling pathways act synergistically to promote angiogenesis. Studies indicate VEGF inhibition leads to increased levels of phosphorylated c-MET, bypassing VEGF-mediated angiogenesis and leading to chemoresistance. We conducted a phase 1 clinical trial with 32 patients with refractory solid tumors to evaluate the safety, pharmacokinetics, and pharmacodynamics of combinations of VEGF-targeting pazopanib and the putative c-MET inhibitor ARQ197 (tivantinib) at 5 dose levels (DLs). Patients either took pazopanib and tivantinib from treatment initiation (escalation phase) or pazopanib alone for 7 days, with paired tumor sampling, prior to starting combination treatment (expansion phase). Hypertension was the most common adverse event. No more than 1 dose limiting toxicity (DLT) occurred at any DL, so the maximum tolerated dose (MTD) was not determined; DL5 (800 mg pazopanib daily and 360 mg tivantinib BID) was used during the expansion phase. Twenty of 31 evaluable patients achieved stable disease lasting up to 22 cycles. Circulating VEGF, VEGFR2, HGF, and c-MET levels were assessed, and only VEGF levels increased. Tumor c-MET levels (total and phosphorylated) were determined in paired biopsies before and after 7 days of pazopanib treatment. Total intact c-MET decreased in 6 of 7 biopsy pairs, in contrast to previously reported c-MET elevation in response to VEGF inhibition. These results are discussed in the context of our previously reported analysis of epithelial-mesenchymal transition in these tumors.

show abstract

Pharmacodynamic Response of the MET/HGF Receptor to Small-Molecule Tyrosine Kinase Inhibitors Examined with Validated, Fit-for-Clinic Immunoassays

Cited by 31 publications

References 52 publications

Effective implementation of novel MET pharmacodynamic assays in translational studies

Effective implementation of novel MET pharmacodynamic assays in translational studies

Met Signaling in Carcinogenesis

Combination therapy with pazopanib and tivantinib modulates VEGF and c-MET levels in refractory advanced solid tumors

Contact Info

Product

Resources

About