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Cancer-related extracellular matrix signatures and related methods and products

a technology of extracellular matrix and signature, applied in the direction of material analysis, instruments, biological material analysis, etc., can solve the problems of limited use of information to identify the role of extracellular matrix (ecm) proteins in disease, difficult to determine the composition of the information, and illness and mortality

Active Publication Date: 2017-11-30
MASSACHUSETTS INST OF TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite extensive study, cancer, and particularly the process of metastasis, remain major causes of illness and mortality.
Large data sets have been generated using genomic and proteomic methods, but the use of that information to identify the role of extracellular matrix (ECM) proteins in disease has been limited.
This is because the ECM is insoluble and crosslinked and its composition has been difficult to determine.

Method used

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[0154]Methods

[0155]Tissue Preparation and ECM Protein Enrichment—Sequential extractions of frozen samples of tissues or tumors were performed using the CNMCS (Cytosol / Nucleus / Membrane / Cytoskeleton) Compartmental Protein Extraction kit (Cytomol, Union City, CA) according to manufacturer's instructions. In brief, frozen tissues (150-200 mg) or tumors (200-j400 mg) were homogenized and extracted sequentially to remove (1) cytosolic proteins (2), nuclear proteins (3), membrane proteins (4), and cytoskeletal proteins leaving a final insoluble fraction enriched for ECM proteins. Fractions were separated on SDS-polyacrylamide gradient gels, transferred to nitrocellulose membranes and probed with antibodies to proteins characteristic of different subcellular compartments.

[0156]Mass Spectrometry—ECM-enriched fractions were solubilized in urea, disulfide bonds reduced and alkylated, and proteins digested with PNGaseF, Lys-C, and trypsin. Solutions that began cloudy upon initial reconstitution...

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Abstract

Proteomic methods for identifying cancer related proteins and related products and kits are provided. The cancer specific proteins are extracellular matrix proteins that are associated with various aspects of cancer. Panels or signature sets of proteins useful in the detection, diagnosis and treatment of cancers as well as monitoring therapeutic progress in a cancer patient are provided herein along with methods for their detection and for their use in targeting imaging and / or therapeutic agents to the tumors via binding to the specified proteins. The proteins were identified using proteomics analyses of tissue samples taken from cancer patients. In certain aspects the proteins are particularly useful in colon cancer patients.

Description

RELATED APPLICATIONS[0001]This application is a divisional of U.S. application Ser. No. 14 / 389,080, filed on Sep. 29, 2014, which is a national staged filing under U.S.C. §371 of PCT International application PCT / US2013 / 031492, entitled “CANCER-RELATED EXTRACELLULAR MATRIX SIGNATURES AND RELATED METHODS AND PRODUCTS,” filed on Mar. 14, 2013, which claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application Ser. No. 61 / 616,981, entitled “EXTRA CELLULAR MATRIX PROTEINS AND CANCER” filed on Mar. 28, 2012, AND to U.S. Provisional Application Ser. No. 61 / 616,987 entitled “CANCER SIGNATURES AND RELATED METHODS AND PRODUCTS” filed on Mar. 28, 2012 which are herein incorporated by reference in their entirety.FEDERALLY SPONSORED RESEARCH[0002]This invention was made with Government support under NIH U54-CA126515 and U54-CA163109. The Government has certain rights in this invention.FIELD OF THE INVENTION[0003]The invention relates in some aspects to application of recently develop...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/574A61K49/00A61K47/68A61K51/10
CPCG01N33/57492G01N33/57415A61K51/1051A61K49/0002A61K47/6855G01N2333/705A61K51/1063A61K51/1093
Inventor HYNES, RICHARD O.NABA, ALEXANDRACLAUSER, KARLCARR, STEVEN A.TANABE, KENNETH
Owner MASSACHUSETTS INST OF TECH