An urgent clinical goal is to identify molecular networks associated with subpopulations of cancer patients who may respond individually to molecular targeted inhibitors. Current molecular targeted therapeutics is directed at protein kinases and/or their phosphorylated substrates. Therefore, measurement of this new class of phosphoprotein signal pathway epitopes in tumor biopsy samples is crucial for individualizing molecular targeted therapies. Phosphoprotein antigen epitopes are not adequately preserved by formalin fixation and paraffin embedding, and freezing of tissue is very expensive and compromises diagnostic accuracy. We propose the advanced development and clinical validation of an innovative and transformative technology for preserving tissue phosphoproteins and diagnostic histomorphology for clinical cancer molecular profiling. Applying knowledge gained under an NIH R21 funded study, we created a novel tissue preservation chemistry that stabilizes all classes of phosphoproteins, is compatible with paraffin embedding, while maintaining complete diagnostic histomorphology, and fully preserving critical diagnostic immunohistology (IHC) antigens including Estrogen Receptor, Progesterone Receptor, HER2, and Ki-67. These IHC antigens are not preserved by special research fixatives used for tissue RNA preservation. Our new non-formalin tissue preservative, termed Biomarker and Histology Preservative (BHP) can be seamlessly introduced into the current community hospital clinical diagnostic workflow with no additional steps or equipment. At the time of procurement, tissue can be immersed directly in the new fixative and processed into a paraffin block for routine diagnosis, obviating the need for costly freezing during shipping or storage. BHP offers the potential for substantial improvements over conventional formalin fixation. In the present application we propose the blinded clinical validation of our novel preservation chemistry in community hospital settings, utilizing a team of international pathologists for validation. The goal of the project is one-step paraffin block stabilization of all classes of cellular phosphoproteins, diagnostic histomorphology, and diagnostic immunohistochemistry antigens, while at the same time maintaining full diagnostic morphology equivalent or superior to standard formalin fixation. We will collect fresh surgical tissue, under informed consent, covering a broad variety of organs and cancer histology to develop an archive of 150 cases of matched paraffin and frozen specimens. We will measure 100 validated phosphoprotein epitopes spanning membrane, cytoplasmic and nuclear compartments from extracted paraffin sections using Reverse Phase Protein Microarray (RPMA) and Laser Capture Microdissection (LCM) technology. Following objective independent validation by diagnostic pathologists, this transformative technology will be ready for widespread clinical and research use. Adoption of the technology would mean that only one diagnostic paraffin block could be used for all classes of molecular profiling rather than the current requirement for multiple blocks. This would increase diagnostic accuracy while substantially reducing costs.

Public Health Relevance

Our technology for preserving proteins, at the time of collection, will enable analysis of molecular targeted kinase inhibitors for individualized therapy, while considerably reducing healthcare costs, and increasing diagnostic accuracy. Our new non-formalin tissue preservative can be seamlessly introduced into the current community hospital clinical diagnostic workflow with no additional steps or equipment.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants Phase II (R33)
Project #
5R33CA157403-02
Application #
8311650
Study Section
Special Emphasis Panel (ZCA1-SRLB-5 (J1))
Program Officer
Chuaqui, Rodrigo F
Project Start
2011-08-15
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
2
Fiscal Year
2012
Total Cost
$309,458
Indirect Cost
$73,454
Name
George Mason University
Department
Other Basic Sciences
Type
Schools of Arts and Sciences
DUNS #
077817450
City
Fairfax
State
VA
Country
United States
Zip Code
22030
Gallagher, Rosa I; Espina, Virginia (2014) Reverse phase protein arrays: mapping the path towards personalized medicine. Mol Diagn Ther 18:619-30
Chiechi, Antonella; Novello, Chiara; Magagnoli, Giovanna et al. (2013) Elevated TNFR1 and serotonin in bone metastasis are correlated with poor survival following bone metastasis diagnosis for both carcinoma and sarcoma primary tumors. Clin Cancer Res 19:2473-85
Holmes, Frankie Ann; Espina, Virginia; Liotta, Lance A et al. (2013) Pathologic complete response after preoperative anti-HER2 therapy correlates with alterations in PTEN, FOXO, phosphorylated Stat5, and autophagy protein signaling. BMC Res Notes 6:507
Golubeva, Yelena; Salcedo, Rosalba; Mueller, Claudius et al. (2013) Laser capture microdissection for protein and NanoString RNA analysis. Methods Mol Biol 931:213-57
Chiechi, Antonella; Mueller, Claudius; Boehm, Kevin M et al. (2012) Improved data normalization methods for reverse phase protein microarray analysis of complex biological samples. Biotechniques 0:1-7
Liotta, Lance A; Petricoin 3rd, Emanuel F (2012) -Omics and cancer biomarkers: link to the biological truth or bear the consequences. Cancer Epidemiol Biomarkers Prev 21:1229-35
Liotta, Lance A; Petricoin, Emanuel (2011) Cancer biomarkers: closer to delivering on their promise. Cancer Cell 20:279-80
Longo, Caterina; Gambara, Guido; Espina, Virginia et al. (2011) A novel biomarker harvesting nanotechnology identifies Bak as a candidate melanoma biomarker in serum. Exp Dermatol 20:29-34
Mueller, Claudius; Edmiston, Kirsten H; Carpenter, Calvin et al. (2011) One-step preservation of phosphoproteins and tissue morphology at room temperature for diagnostic and research specimens. PLoS One 6:e23780
Espina, Virginia; Mueller, Claudius; Liotta, Lance A (2011) Phosphoprotein stability in clinical tissue and its relevance for reverse phase protein microarray technology. Methods Mol Biol 785:23-43

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