Dr. Margaret E. Black
Research & Interests
Exploiting nucleoside metabolizing enzymes for novel cancer therapies is a major focus of my lab's research. We are fundamentally interested in the structure to function relationship of enzymes that catalyze the formation of precursors for DNA and RNA and, most notably, also convert nucleoside analogs or prodrugs to cytotoxic compounds. In a process known as suicide gene therapy, a nucleoside metabolizing gene is delivered to a cancer cell followed by administration of the nontoxic prodrug. The gene product (enzyme) converts the prodrug to a cytotoxin, thereby leading to death of the cancer cell. Our approach employs several molecular strategies to generate enzyme variants with improved activities towards the nontoxic prodrugs for use in suicide gene therapy. Biochemical evaluation along with in vitro and in vivo analysis of derived mutants that display enhanced prodrug activation allows the identification of mutants for use in gene therapy for the treatment of cancer. Molecular modeling of the active site of enzymes and their variants has provided further insights on how the enzyme might be altered to improve prodrug activation and therefore enhance tumor ablation for a safer and more effective cancer cure.
Recently we have extended the application of these novel suicide genes beyond gene therapy by incorporating targeting motifs for ‘Targeted Enzyme Therapy’. In this approach the targeting motif directs the suicide enzyme to bind to well established biomarkers found on the cancer cell surface. Following prodrug administration, the cytotoxic compound generated by the action of the suicide enzyme enters the cell and elicits cell death.
Teaching Expertise: I am course director for MBios 494 (Senior Projects) in Fall and for MBios/ChE 574 a graduate course called Protein Biotechnology offered every other year in Spring semester. In addition, in Fall I teach prokaryotic gene regulation in MBios 503 (Advanced Molecular Biology).
- L. Dore-Savard, Z. Chen, P. T. Winnard, B. Krishnamachary, V. Raman, M. E. Black and Z. M. Bhujwalla. (2016) Delayed Progression of Lung Metastases Following Delivery of a Prodrug Activating Enzyme. Clinical and Experimental Metastasis. Submitted.
H. Ruan and M. E. Black. (2016) Zebularine-Resistant Human Cytidine Deaminase Mutants for Optimal Chemoprotection of Hematopoietic Stem Cells. J. Genet. Syndr. Gene Ther. 7:4.
C. Souza, N. Villarino, K. Farnsworth and M. E. Black. (2016) Enhanced Cytotoxicity of Bleomycin, Cisplatin, and Carboplatin on Equine Sarcoid Cells following Electroporation-Mediated Delivery In Vitro. J. Vet. Pharmacol. Ther., ePub June 11, 2016.
H. Ruan, S. Qui, B. Beard and M. E. Black. (2016) Chemoprotection of Hematopoietic Stem Cells via Creation of Zebularine Resistant Human Cytidine Deaminase Mutants. Protein Engineering, Selection and Design. ePub May 8, 2016.
S. E. Martin, T. Ganguly, G. R. Munske, M. D. Fulton, M. R. Hopkins, C. E. Berkman and M. E. Black. (2014) Development of Inhibitor-Directed Enzyme Prodrug Therapy (IDEPT) for Prostate Cancer. Bioconjugate Chemistry, 25:1752-60.
A. Ardiani, A. J. Johnson, H. Ruan, M. Sanchez-Bonilla, K. Serve and M. E. Black. (2012) Enzymes To Die For: Exploiting Nucleotide Metabolizing Enzymes for Cancer Gene Therapy. Current Gene Therapy, 12(2):77-91. (Peer-reviewed review).
A. J. Johnson, M. N. Brown and M. E. Black. (2011) Evaluation of a UCMK/dCK Fusion Enzyme for Gemcitabine-Mediated Cytotoxicity. Biochem. Biophys. Res. Comm., 416:199-204.
A. J. Johnson*, A. Ardiani*, M. Sanchez-Bonilla and M. E. Black. (2011) Comparative Analysis of Enzyme and Pathway Engineering Strategies for 5FC-Mediated Suicide Gene Therapy Applications. * - joint first authors. Cancer Gene Therapy, 18:533-542.
K. M. Serve, J. Darnell, J. K. Takemoto, N. M. Davies and M. E. Black. (2010) Validation of an Isocratic HPLC Method to Detect 2-Fluoro-β-Alanine for the Analysis of In Vitro Dihydropyrimidine Dehydrogenase Activity. J. Chromot. B, 878:1889-1892.
K.M. Serve, J. A. Yanez, C. M. Remsberg, N. M. Davies and M. E. Black. (2010) Development and Validation of a Rapid and Sensitive HPLC Method for Detection of 5-Fluorocytosine and Its Metabolites. Biomed. Chromat., 24:556-561.
A. Ardiani, M. Sanchez-Bonilla and M E. Black. (2010) Fusion Enzymes Containing HSV-1 Thymidine Kinase Mutants and Guanylate Kinase Enhance Prodrug Sensitivity In Vitro and In Vivo. Cancer Gene Therapy, 17:86-96.
M. Fuchita*, A. Ardiani*, L. Zhao, B. L. Stoddard and M. E. Black. (2009) Engineering Bacterial Cytosine Deaminase for Efficient 5FU Production for Cancer Gene Therapy. * -joint first authors. Cancer Research, 69:4791-4799.
A. Ardiani, A. Goyke and M. E. Black. (2009) Mutations at Serine 37 in Mouse Guanylate Kinase Confer Resistance to 6-Thioguanine. Protein Engineering, Selection and Design, 22:225-232.
M. Johnson, B. D. W. Karanikolas, S. J. Priceman, R. Powell, M. E. Black, C. Wu, J. Czernin, H. Huang and L. Wu. (2009) Titration of Variant HSV1-tk Gene Expression to Determine the Sensitivity of 18F-FHBG PET Imaging in a Prostate Tumor. J. Nuclear Med., 50:757-764.
L. N. Kaliberova, D. D. Manna, V. Krendelchtchikova, M. E. Black, D. J. Buchsbaum and S. A. Kaliberov. (2008) Molecular Chemotherapy of Pancreatic Cancer Using Novel Mutant Bacterial Cytosine Deaminase Gene. Molec. Cancer Therapy, 7:2845-2854.
C. L. Willmon, D. Sussman and M. E. Black. (2008) The Role of Herpes Simplex Virus-1 Thymidine Kinase Alanine 168 in Substrate Specificity. The Open Biochem. J., 2:60-66.
T. S. Stolworthy*, A. Korkegian*, C. L. Willmon, A. Ardiani, J. Cundiff, B. L. Stoddard and M. E. Black. (2008) Yeast Cytosine Deaminase Mutants with Increased Thermostability Impart Sensitivity to 5-Fluorocytosine. J. Mol. Biol., 377:854-869. * -joint first authors.
S. A. Kaliberov, J. M. Markert, V. Krendelchtchikova, D. D. Manna, J. C. Sellers, L. N. Kaliberova, M. E. Black and D. J. Buchsbaum. (2007) Gene Directed/Enzyme Prodrug Therapy of Human Glioma Using Mutant Escherichia coli Cytosine Deaminase. Gene Therapy, 14:1111-1119.
C. L. Willmon, E. Krabbenhoft and M. E. Black. (2006) A Guanylate Kinase/HSV-1 Thymidine Kinase Fusion Protein Enhances Prodrug Mediated Cell Killing. Gene Therapy, 13:1309-1312.
A. Korkegian, M. E. Black, D. Baker and B. L. Stoddard. (2005) Computational Thermostabilization of an Enzyme. Science, 308:857-860.