Abnormalities in phosphosignaling networks in cancers are often the direct or indirect targets of anti-tumor agents. Thus it is critical that we are able to reliably quantify phosphoproteins in cancer biopsies to establish mechanism of action, to evaluate pharmacodynamic responses (to inform dose and scheduling), and to monitor compensatory pathways that influence efficacy and toxicity. Prior studies have shown that a significant subset of the cancer phosphoproteome is highly susceptible to preanalytical variables, such as ischemic time prior to fixation or freezing. Thus, it is critical that the collection and processing of cancer biopsies be conducted using a strict SOP that minimizes preanalytical variations. For extraction-based methods (e.g. mass spectrometry, ELISA, Western blotting), the current gold standard is to flash freeze tumor samples in liquid nitrogen, followed by preparation of protein lysates in denaturing conditions in the presence of kinase/phosphatase inhibitor cocktails. However, a major limitation is that in many clinical settings liquid nitrogen is not readily available, and neither the personnel nor the infrastructure is generally available to rapidly process the tumor samples. As a result, tumors are often subjected to prolonged ischemia and/or chemical fixatives, altering the phosphoproteome and thereby compromising the biospecimen?s integrity such that it may no longer reflect the true in vivo state of the tumor. We propose to develop an affordable, convenient, single-use device for rapidly freezing tumor biopsies in clinical settings to preserve the labile phosphoproteome for molecular analyses.

Public Health Relevance

Abnormalities in phosphosignaling networks in cancers are often the direct or indirect targets of anti-tumor agents. Thus it is critical that we are able to reliably quantify phosphoproteins in cancer biopsies to establish mechanism of action, to evaluate pharmacodynamic responses (to inform dose and scheduling), and to monitor compensatory pathways that influence efficacy and toxicity. We propose to develop an affordable, convenient, single-use device for rapidly freezing tumor biopsies at point of care in clinical settings to preserve the labile phosphoproteome for molecular analyses, which should have a large impact on our understanding of cancer biology and the pharmacodynamics of potential novel cancer therapies.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21CA225507-02
Application #
9706812
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Agrawal, Lokesh
Project Start
2018-06-01
Project End
2020-05-31
Budget Start
2019-06-01
Budget End
2020-05-31
Support Year
2
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
078200995
City
Seattle
State
WA
Country
United States
Zip Code
98109