Dr. Eric B. Kmiec

Professor

Contact

Kmiec

Lab: 270 DBI

Mailing address:
Delaware Biotechnology Institute
Delaware Technology Park
15 Innovation Way
Newark DE, 19711

Phone: (302) 831-3420
Fax: (302) 831-3427
E-mail: ekmiec@udel.edu
Web: Kmiec Lab Homepage

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Education

B.A.: Rutgers University
M.S.: Southern Illinois University
Ph.D.: University of Florida School of Medicine

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Research Interests

The research focus of this laboratory, centers on the development of nucleic acid therapeutics particularly on molecules that can either direct genetic changes in the chromosomes of mammalian cells and animal models or adopt unique conformations to block mutant protein activity. We use biochemical and genetic assay systems to study the mechanism by which these changes occur and how the cell regulates these genetic manipulations. Presently, the vector of choice for gene repair is a single-stranded DNA oligonucleotide that directs the reversal of single base mutations by engaging the endogenous DNA repair and recombination activities of the cell. The repair of single base mutations by these oligonucleotides is being tested as a gene therapy approach for several inherited disorders including muscular dystrophy and spinal muscular atrophy. Single-stranded vectors are also being tested for efficacy in animal models of human diseases through multiple collaborations at major medical centers. These targets include Criglar-Najjar Disease and osteogenesis imperfecta.

The laboratory continues to explore the molecular mechanism by which single base changes are made. In 2005, the Kmiec group proposed a model in which the oligonucleotide is incorporated into a growing strand of DNA during replication. Independent laboratories have now confirmed that his mechanism likely accounts for the initial phases of gene repair although others are still possible. Thus, the laboratory is engaged in developing novel techniques that can alter the rate of DNA replication in order to increase the efficiency and with which gene repair occurs. Importantly, full chromosomal replication need not occur for the oligo to direct the change. The elevation of recombination and replication proteins in the cell suffices to increase the frequency of gene repair.

Single-stranded oligonucleotides are also being used as aptamers to inhibit the pathogenecity of Huntington's Disease (HD). Molecules of monomeric guanosine sequence capable of adopting a G-quartet conformation have been found to efficiently block HD micro-aggregation processes. This small nontoxic structure appears to bind to the mis-folded HD protein as well as the mRNA and prevent nucleation of aggregation events. Our lab is utilizing both biochemical and cell-based assays to identify variant molecules that exhibit the highest level of efficacy and to understand the mechanism of this unique activity in detail. G-quartets are also being used to induce an apoptosis event in esophageal cancer cells. These molecules are displaying unique properties that allow them to distinguish between normal and malignant cell types. The mechanism by which this occurs is being investigated at the cell and animal models.

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Current Projects

• Mechanism of oligonucleotide-directed gene repair - Using a genetic readout system and cell free extracts from human and plant cells, the molecular and biochemical events underlying gene repair are being studied in yeast, bacteria and mammalian cells.
• Development of DNA base therapeutics for Huntington's Disease - A cell-based assay system is being employed to explore ways to prevent the aggregation activity of the Huntington protein.
• Gene therapy for inherited disorders - Primary patient cells are being used to test the efficacy of gene editing for Spinal Muscular Atrophy (SMA) and Muscular Atrophy.
• Collaborative cancer research - Introduction of oligonucleotides that form G-quartet conformations, to induce apoptosis in esophageal cancer cells.

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Teaching

• BISC 403* - Genetic and Evolutionary Biology
• BISC 600 - Molecular Medicine and Biotechnology

*Course web site available through MyCourses

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Research Group

• Takayuki Suzuki, Ph.D. - Postdoctoral Researcher (Ph.D., Nagoya University, Japan). Gene editing as a therapy for Muscular Dystrophy, oligonucleotide delivery and increasing gene repair efficiency in mouse models.
• Michael Skogen, M.S. - Research Technician (M.S., University of Delaware). Mechanism of apoptosis in cancer cells by G-rich oligonucleotides.
• Darlise M. DiMatteo, B.A. - Research Associate (B.A,. University of Delaware). Gene editing as a therapy for Spinal Muscular Atrophy (SMA).
• Julia Engstrom, B.S. - Graduate Student (B.S., University of Delaware). Gene targeting and mechanism of gene repair in disease models.
• Carly Falgowksi, B.S. - Research Specialist (B.S., College of William and Mary). Gene correction in Spinal Muscular Atrophy (SMA).
• Kerry Falgowksi, B.S. - Graduate Student (B.S., University of North Carolina at Chapel Hill). C. elegans as a model system for treatment of muscular dystrophy.
• Hetal Parekh-Olmedo, B.S. - Senior Research Associate/Lab Manager (B.S., Holy Family University). Mechanism and therapeutic applications of oligonucleotides for the treatment of human disease.
• Jennifer Roth, B.S. - Graduate Student (B.S., Susquehanna University). DNA damage pathways and genetic events surrounding the expression of mutant huntingtin protein.
• Tim Schwartz, B.S. - Graduate Student (B.S., Gettysburg College). Targeted repair in mammalian cells and cancer gene therapy.
• Sarah Yerkes, B.S. - Graduate Student (B.S., University of Delaware). Use of oligonucleotides as a therapeutic in Huntington's Disease and study of the interaction of G-rich oligonucleotides with the huntingtin protein.
• Stephanie Callahan - Undergraduate Student. Gene editing as a treatment for Spinal Muscular Atrophy (SMA).
• Melissa Warriner - Undergraduate Student. Microarray analysis of treated esophageal cancer cells.

The Laboratory of Applied Genomics

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Selected Publications

Ferrara L, Engstrom J, Schwartz T, Parekh-Olmedo H and Kmiec EB (2007). Recovery of corrected cells after initiation of the targeted gene repair reaction. DNA Repair (Amst). Jun 8; [Epub ahead of print].

Engstrom J and Kmiec EB (2007). Manipulation of S phase progression can counteract cell cycle arrest of corrected cells during the oligonucleotide-directed gene repair reaction. BMC Molecular Biology 8, 9.

Maguire K and Kmiec EB (2007). Multiple roles for MSH2 in the repair of a deletion mutation directed by modified single-stranded oligonucleotides. Gene 386(1-2), 107-114.

Schwartz T and Kmiec EB (2007). Reduction of gene repair by selenomethionine with the use of single-stranded oligonucleotides. BMC Molecular Biology 8, 7.

Skogen M, Roth J, Parekh-Olmedo H and Kmiec EB (2006). Short G-rich oligonucleotides as a therapeutic for Huntington’s Disease (HD). BMC Neuroscience 7, 65.

Ferrara L and Kmiec EB (2006). Targeted gene repair activates Chk1 and Chk2 and stalls replication in corrected cells. DNA Repair (Amst). 5(4), 422-431.

Schwartz T and Kmiec EB (2005). Using methyl methanesulfonate (MMS) to stimulate targeted gene repair activity in mammalian cells. Gene Therapy and Molecular Biology 9, 193-202.

Engstrom J and Kmiec EB (2005). Caffeine elevates and stabilizes gene repair efficiencies in mammalian cells. Gene Therapy and Molecular Biology 9, 445-456.

Drury M, Skogen M and Kmiec EB (2005). A tolerance of DNA heterology in the mammalian targeted gene repair reaction. Oligonucleotides 15(3), 155-171.

Parekh-Olmedo H, Ferrara L and Kmiec EB (2005). Progress and Prospects: targeted gene repair. Gene Therapy 12, 639-646.

Brachman E and Kmiec EB (2005). Gene repair in mammalian cells is stimulated by the elongation of S phase and transient stalling of replication forks. DNA Repair 4, 445-457.

Hu Y, Parekh-Olmedo H, Drury M, Skogen M and Kmiec EB (2005). Reaction parameters of targeted gene repair in mammalian cells. Molecular Biotechnology 29, 197-210.

Drury M and Kmiec EB (2004). Double displacement loops (double d-loops) are a more active template for gene repair activity. Oligonucleotides 14(4), 274-286.

Luciana F, Kmiec EB (2004). Camptothecin enhances the frequency of oligonucleotide-directed gene repair in mammalian cells by inducing DNA damage and activating homologous recombination. Nucleic Acids Research. 32(17), 5239-5248.

Luciana F, Parekh-Olmedo H, Kmiec EB (2004). DNA damage increases the frequency of gene repair in mammalian cells. Experimental Cell Research. 300(1), 170-179.

Brachman E and Kmiec EB (2004). DNA replication impacts repair frequency and transcription strand bias in targeted nucleotide exchange. J. Cell Science. 117(17), 3867-3874.

van Brabant A, Williams JK, Parekh-Olmedo H, and Kmiec EB (2004). Gene editing of a human gene in yeast artificial chromosomes using modified single-stranded DNA and dual targeting. The Pharmacogenomics Journal. 4, 175-183.

Liu L, Katie K. Maguire, and Kmiec EB (2004). Engineering the eukaryotic recombinase Rad51 increases the frequency of gene repair in vivo. Nucl. Acids Res. 32, 2093-2101.

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