#9 - Tissue-based Diagnostic Assay
Title: Ratio Based Biomarkers for the Prediction of Cancer Survival
NIH Reference No.: E-025-2009
Executive Summary
General Description
The inventors have developed a method of analyzing multiple protein biomarkers, assayed in formalin fixed, paraffin embedded tissues that allows the combination of between two and four biomarkers to result in an outcome measure of two categories. The approach has been demonstrated applicable to multiple cancer systems. The approach is based on profiling a cancer based on pathway activation, and can be applied to different biomarkers via different platforms, on current clinical samples
Substantial data of this approach is on Akt, or protein kinase B, which plays a key role in the regulation of important cellular processes including cellular survival, apoptosis, and protein translation. The Akt pathway has been shown to have prognostic significance in a number of cancers and is associated with tumor cell survival, proliferation and invasiveness. The activation of Akt is frequently observed in human cancers and tumor cells. Akt is activated by phosphorylation of Thr308 by PIP3 and at Ser473 by mammalian target of rapamycin (mTOR). The phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a known negative regulator of the Akt signaling pathway which functions as a tumor suppressor gene
Scientific Progress
Recently, NCI inventors have identified several functions of the Akt pathway in certain cancers, such as extrahepatic cholangiocarcinoma (EHCC). Samples from 221 EHCC patients with clinicopathologic data were arrayed into tissue microarrays. Phosphorylated Akt (pAkt), phosphorylated mammalian target of rapamycin (p-mTOR), and PTEN protein expressions were studied with multiplex tissue immunoblotting assay. Expressions of pAkt and p-mTOR were significantly increased in EHCC cases when compared to normal and dysplastic bile duct epithelium (P < 0.05 for both). Decreased PTEN expression (P = 0.004), as well as decreased PTEN/pAkt (P = 0.003) and PTEN/p-mTOR (P = 0.009) expression showed shorter survival by univariate but not by multivariate analysis. The technology has also been demonstrated to function in lung cancer, by means of traditional immunohistochemistry, coupled with image analysis. An algorithm of (p-mTOR/p-AKT) + (p-MAPK/EGFR) = outcome value was developed. In addition, predictive markers were identified in 946 patient gastric cancer samples. From the multivariate analysis of numerous markers, HER2 /HER3 was a stronger predictor of survival, than either HER2 or HER3 alone for gastric cancer patients
This technology is a method for identifying and characterizing biomarkers expressed in a sample that are associated with the presence, the development or progression of cancer. The technology could be made into a kit for use in the clinic. Overall, the inventors have demonstrated that in situ detection of ratios of PTEN/p-AKT ratios, and PTEN/p-mTOR, p-mTOR/p-AKT, and p-MAPK/EGFR as well as the combination of the later two, can predict the survival of cancer patients. These biomarkers may provide useful diagnostic information for cancer patients as well as identify patients appropriate for mTOR analog-based chemotherapy or agents directed against AKT. Ratios of HER2/HER3 also have predictive value. The approach can be applied to different methods of detecting proteins from in situ formalin fixed, paraffin embedded tissue samples
Future Work
Strengths
Weaknesses
Patent Status
U.S. Patent Application No. 13/144,474 filed 13 July 2011 Google Patent
Also filed in Canada, Europe (Validated in Germany, France, and UK), Australia, Japan, Israel, and issued in New Zealand
U.S. Patent Application No. 13/841,176 filed 15 March 2013 PatentStorm
Relevant Publications
JY Chung et al. Clin Cancer Res. 2009 Jan 15;15(2):660-667. (PubMed ID: 19147772)
JY Chung et al. Proteomics 2006 Feb;6(3):767-774. (PubMed ID: 16400680)
JY Chung et al. Cancer Epidemiol Biomarkers Prev. 2006 Jul;15(7):1403-1408. (PubMed ID: 16835344)
Matsuda KM, Chung JY, Hewitt SM. Expert Rev Proteomics. 2010;7(2):227-37. (PubMed ID: 20377389)
Kitano H, et al. The combination of Phospho-ATK, Phospho mTOR, Phospho MAPK, and EGFR predicts survival in non-small cell lung cancer (submitted).
Inventor Bio
Stephen M. Hewitt, M.D., Ph.D.
Stephen M. Hewitt, M.D., Ph.D., is appointed as a Clinical Investigator within the Laboratory of Pathology, Center for Cancer Research, National Cancer Institute at the National Institutes of Health, in Bethesda Maryland. He has served as chief of the Tissue Array Research Program since its inception in 2000, and as chief of the Applied Molecular Pathology Laboratory since its creation in 2008
Dr. Hewitt's research interests are in the development of tissue-based biomarkers for cancer diagnosis, prognosis and prediction of response to therapy. Specific areas of concentration include tissue microarrays, biobanking of pathology specimens, tissue proteomics, whole slide imaging /image analysis, and cancers of the aero-digestive and urogenitial tracts
Stephen received his bachelor's degree in philosophy from the Johns Hopkins University in 1988, and his Ph.D. in genetics from the University of Texas, Graduate School of Biomedical Sciences in 1995, having completed his thesis in the laboratory of Grady Saunders at the University of Texas M.D. Anderson Cancer Center. He completed his M.D. in 1996 at the University of Texas Medical School at Houston, and his residency in Anatomic Pathology within the Laboratory of Pathology at the National Cancer Institute. He is a board certified Anatomic Pathologist and Fellow of the College of American Pathologist and Fellow of the American Society of Clinical Pathology. Dr. Hewitt is a member of the Clinical Laboratory Standards Institute Immunology & Ligand Assay Consensus Committee, having served as co-chair of the Subcommittee on Immunohistochemical Assays. He is a councilor of the Histochemical Society, Program Committee Chair-elect for the Association for Pathology Informatics and a consultant to the Hematology and Pathology Devices Panel, Center for Devices and Radiological Health, Food and Drug Administration. Dr. Hewitt has co-authored more than 200 articles and servers on the editorial board of four peer-reviewed journals
NIH Reference No.: E-025-2009
Executive Summary
- Category: Diagnostic
- Disease Focus: Cancer broadly
- Basis of Invention: Protein
- How it works: Combines two to four biomarkers to predict a patient’s survival and determine treatment course
- Patent Status: Pending in U.S., Canada, Europe, Australia, Japan, Israel, and New Zealand
- Lead Inventor: Stephen M. Hewitt, M.D., Ph.D.
- Development Stage: The biomarkers have been evaluated in 221 patients with extrahepatic cholangiocarcinoma (EHCC), in 220 lung cancer patients, in 946 patients with gastric cancer, as well as in cervical and kidney cancers.
- Novelty: Novel biomarkers that predict patient survival
- Clinical Application: Prognostic / predictive biomarkers for survival and prediction of response to targeted therapies. Identification of patients who may benefit from mTOR analog-based chemotherapy or agents directed against Akt
General Description
The inventors have developed a method of analyzing multiple protein biomarkers, assayed in formalin fixed, paraffin embedded tissues that allows the combination of between two and four biomarkers to result in an outcome measure of two categories. The approach has been demonstrated applicable to multiple cancer systems. The approach is based on profiling a cancer based on pathway activation, and can be applied to different biomarkers via different platforms, on current clinical samples
Substantial data of this approach is on Akt, or protein kinase B, which plays a key role in the regulation of important cellular processes including cellular survival, apoptosis, and protein translation. The Akt pathway has been shown to have prognostic significance in a number of cancers and is associated with tumor cell survival, proliferation and invasiveness. The activation of Akt is frequently observed in human cancers and tumor cells. Akt is activated by phosphorylation of Thr308 by PIP3 and at Ser473 by mammalian target of rapamycin (mTOR). The phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is a known negative regulator of the Akt signaling pathway which functions as a tumor suppressor gene
Scientific Progress
Recently, NCI inventors have identified several functions of the Akt pathway in certain cancers, such as extrahepatic cholangiocarcinoma (EHCC). Samples from 221 EHCC patients with clinicopathologic data were arrayed into tissue microarrays. Phosphorylated Akt (pAkt), phosphorylated mammalian target of rapamycin (p-mTOR), and PTEN protein expressions were studied with multiplex tissue immunoblotting assay. Expressions of pAkt and p-mTOR were significantly increased in EHCC cases when compared to normal and dysplastic bile duct epithelium (P < 0.05 for both). Decreased PTEN expression (P = 0.004), as well as decreased PTEN/pAkt (P = 0.003) and PTEN/p-mTOR (P = 0.009) expression showed shorter survival by univariate but not by multivariate analysis. The technology has also been demonstrated to function in lung cancer, by means of traditional immunohistochemistry, coupled with image analysis. An algorithm of (p-mTOR/p-AKT) + (p-MAPK/EGFR) = outcome value was developed. In addition, predictive markers were identified in 946 patient gastric cancer samples. From the multivariate analysis of numerous markers, HER2 /HER3 was a stronger predictor of survival, than either HER2 or HER3 alone for gastric cancer patients
This technology is a method for identifying and characterizing biomarkers expressed in a sample that are associated with the presence, the development or progression of cancer. The technology could be made into a kit for use in the clinic. Overall, the inventors have demonstrated that in situ detection of ratios of PTEN/p-AKT ratios, and PTEN/p-mTOR, p-mTOR/p-AKT, and p-MAPK/EGFR as well as the combination of the later two, can predict the survival of cancer patients. These biomarkers may provide useful diagnostic information for cancer patients as well as identify patients appropriate for mTOR analog-based chemotherapy or agents directed against AKT. Ratios of HER2/HER3 also have predictive value. The approach can be applied to different methods of detecting proteins from in situ formalin fixed, paraffin embedded tissue samples
Future Work
- Validation of described markers
- Expansion to additional tumor types
- Expansion to encompass additional pathways
Strengths
- Further along in development as it is already tested in patient samples
- Statistically significant findings
- Patent filed internationally allowing for market analysis in multiple regions
Weaknesses
- Will need evaluation/validation in additional cancer types, including breast cancer, to determine the broader applicability of the biomarkers
Patent Status
U.S. Patent Application No. 13/144,474 filed 13 July 2011 Google Patent
Also filed in Canada, Europe (Validated in Germany, France, and UK), Australia, Japan, Israel, and issued in New Zealand
U.S. Patent Application No. 13/841,176 filed 15 March 2013 PatentStorm
Relevant Publications
JY Chung et al. Clin Cancer Res. 2009 Jan 15;15(2):660-667. (PubMed ID: 19147772)
JY Chung et al. Proteomics 2006 Feb;6(3):767-774. (PubMed ID: 16400680)
JY Chung et al. Cancer Epidemiol Biomarkers Prev. 2006 Jul;15(7):1403-1408. (PubMed ID: 16835344)
Matsuda KM, Chung JY, Hewitt SM. Expert Rev Proteomics. 2010;7(2):227-37. (PubMed ID: 20377389)
Kitano H, et al. The combination of Phospho-ATK, Phospho mTOR, Phospho MAPK, and EGFR predicts survival in non-small cell lung cancer (submitted).
Inventor Bio
Stephen M. Hewitt, M.D., Ph.D.
Stephen M. Hewitt, M.D., Ph.D., is appointed as a Clinical Investigator within the Laboratory of Pathology, Center for Cancer Research, National Cancer Institute at the National Institutes of Health, in Bethesda Maryland. He has served as chief of the Tissue Array Research Program since its inception in 2000, and as chief of the Applied Molecular Pathology Laboratory since its creation in 2008
Dr. Hewitt's research interests are in the development of tissue-based biomarkers for cancer diagnosis, prognosis and prediction of response to therapy. Specific areas of concentration include tissue microarrays, biobanking of pathology specimens, tissue proteomics, whole slide imaging /image analysis, and cancers of the aero-digestive and urogenitial tracts
Stephen received his bachelor's degree in philosophy from the Johns Hopkins University in 1988, and his Ph.D. in genetics from the University of Texas, Graduate School of Biomedical Sciences in 1995, having completed his thesis in the laboratory of Grady Saunders at the University of Texas M.D. Anderson Cancer Center. He completed his M.D. in 1996 at the University of Texas Medical School at Houston, and his residency in Anatomic Pathology within the Laboratory of Pathology at the National Cancer Institute. He is a board certified Anatomic Pathologist and Fellow of the College of American Pathologist and Fellow of the American Society of Clinical Pathology. Dr. Hewitt is a member of the Clinical Laboratory Standards Institute Immunology & Ligand Assay Consensus Committee, having served as co-chair of the Subcommittee on Immunohistochemical Assays. He is a councilor of the Histochemical Society, Program Committee Chair-elect for the Association for Pathology Informatics and a consultant to the Hematology and Pathology Devices Panel, Center for Devices and Radiological Health, Food and Drug Administration. Dr. Hewitt has co-authored more than 200 articles and servers on the editorial board of four peer-reviewed journals