Principal Investigator

Peggy M
Angel
Awardee Organization

Medical University Of South Carolina
United States

Fiscal Year
2020
Activity Code
R21
Project End Date

Enzymatic Tools for 2D Tissue Localized and Deeper Proteomic Sequencing of Cancer Stromal Proteins

Determining the role of stromal proteins in cancer biology has remained difficult due to a lack of tools. Current tools are qualitative and cannot report on protein sequence variation that is critical to understanding how stromal proteins help or impede cancer growth and metastasis. Our previous work has shown that bacterial collagenases can be used to produce stromal proteomes detected by mass spectrometry as 2D distributions on cancer tissues. We still lack tools that can focus down on target proteins for detailed studies of sequence variation and posttranslational modification. We propose using matrix metalloproteinases (MMPs) as enzymatic tools for targeted detection of stromal protein sequences in the cancer tissue microenvironment. Aim 1, we will develop and optimize a suite of MMPs for analysis of stromal sequences in tumor tissue sections. We will produce and optimize recombinant MMPs specifically for use on 2D solid substrates of tissue sections. We will integrate sequence information with current qualitative tools, expanding capabilities for stromal protein sequence detection down to a single cell niche. In Aim 2, we will focus on developing standards for quantitative detection of target stromal proteins from the tissue microenvironment. We will test the developed technology against a small but relevant cohort of emergent hepatocellular carcinoma from cirrhosis. Our research team has a unique combination of expertise for the production, development, and future commercialization of MMPs. The proposal is transformative since it will allow detection of stromal protein sequence variation from fixed clinical specimens which is impossible with conventional methods. A long-term objective is to significantly improve the community's ability to target stromal proteins within the tumor tissue microenvironment for use in basic research, directing therapies, or prognostic or diagnostic biomarkers.

Publications

  • Angel PM, Spruill L, Jefferson M, Bethard JR, Ball LE, Hughes-Halbert C, Drake RR. Zonal regulation of collagen-type proteins and posttranslational modifications in prostatic benign and cancer tissues by imaging mass spectrometry. The Prostate. 2020 Sep;80(13):1071-1086. Epub 2020 Jul 20. PMID: 32687633
  • Angel PM, Rujchanarong D, Pippin S, Spruill L, Drake R. Mass Spectrometry Imaging of Fibroblasts: Promise and Challenge. Expert review of proteomics. 2021 Jun;18(6):423-436. Epub 2021 Jul 24. PMID: 34129411
  • Clift CL, McLaughlin S, Muñoz M, Suuronen EJ, Rotstein BH, Mehta AS, Drake RR, Alarcon EI, Angel PM. Evaluation of Therapeutic Collagen-Based Biomaterials in the Infarcted Mouse Heart by Extracellular Matrix Targeted MALDI Imaging Mass Spectrometry. Journal of the American Society for Mass Spectrometry. 2021 Dec 1;32(12):2746-2754. Epub 2021 Oct 29. PMID: 34713699
  • Clift CL, Drake RR, Mehta A, Angel PM. Multiplexed imaging mass spectrometry of the extracellular matrix using serial enzyme digests from formalin-fixed paraffin-embedded tissue sections. Analytical and bioanalytical chemistry. 2021 Apr;413(10):2709-2719. Epub 2020 Nov 18. PMID: 33206215
  • Angel PM, Drake RR, Park Y, Clift CL, West C, Berkhiser S, Hardiman G, Mehta AS, Bichell DP, Su YR. Spatial N-glycomics of the human aortic valve in development and pediatric endstage congenital aortic valve stenosis. Journal of molecular and cellular cardiology. 2021 May;154:6-20. Epub 2021 Jan 29. PMID: 33516683
  • Clift CL, Su YR, Bichell D, Jensen Smith HC, Bethard JR, Norris-Caneda K, Comte-Walters S, Ball LE, Hollingsworth MA, Mehta AS, Drake RR, Angel PM. Collagen fiber regulation in human pediatric aortic valve development and disease. Scientific reports. 2021 May 7;11(1):9751. PMID: 33963260
  • Rujchanarong D, Lefler J, Saunders JE, Pippin S, Spruill L, Bethard JR, Ball LE, Mehta AS, Drake RR, Ostrowski MC, Angel PM. Defining the Tumor Microenvironment by Integration of Immunohistochemistry and Extracellular Matrix Targeted Imaging Mass Spectrometry. Cancers. 2021 Sep 1;13. (17). PMID: 34503228