David R Corey
University Of Texas Sw Med Ctr/dallas
Project start date: 2005-02-01
Project end date: 2013-01-31
Sponsored Links Excellgen http://Excellgen.com
RECOGNITION OF CHROMOSOMAL DNA BY CHEMICALLY MODIFIED OLIGONUCLEOTIDES
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas, Dallas, Tx 75390-9105
Grant 5R01GM073042-06 from National Institute Of General Medical Sciences
Abstract: Synthetic agents that recognize specific sequences within chromosomal DNA are a promising strategy for controlling gene expression. During the previous funding period we described duplex RNAs, peptide nucleic acids (PNAs), PNA-peptides, and locked nucleic acids (LNAs) that target gene promoters and modulate expression. LNA bases contain a modified ribose with a methylene linkage between the 2´-oxygen and 4´-carbon and increase melting temperature values (Tm´s) by as much as 100C per substitution. We focus this proposal on LNAs because LNAs have been the simplest and most effective tools for direct recognition of chromosomal DNA. Objectives. Our objective for the next funding period is to understand the potential of chromosome- targeted LNAs as research tools and leads for clinical development. Research Design. We will investigate the mechanism, optimization and applications of agLNAs. In Aim 1 we will investigate the mechanism of agLNA-mediated recognition of nucleic acid targets and subsequent inhibition of gene expression inside cells. The environment surrounding gene promoters is a complex mix of RNA transcripts, proteins, and chromosomal DNA. Understanding the mechanism of recognition will help guide use of agLNAs for new applications and provide important basic insights into how synthetic oligomers affect cellular processes at gene promoters. In Aim 2, we will characterize cellular uptake, improve delivery methods, and test chemically modified LNAs. The goal for this Aim is to identify the most straightforward and efficient combination of delivery protocol and chemical structure for achieving optimal inhibition with antigene oligonucleotides. This information will assist researchers hoping to use agLNAs and help guide the design of agLNAs for therapeutic applications. In Aim 3 we propose novel strategies to use agLNAs to control cellular processes. Applications include use of agLNAs to probe accessibility of sequences within chromosomal DNA, targeting agLNAs to genes with bidirectional promoters (10 % of human genes), using agLNAs to manipulate expression of genes that are important biomedical targets, and synthesis and testing of agLNA-peptide conjugates designed to recruit transcription factors to gene promoters and activate gene expression. These experiments were chosen for their potential to expand the range of applications for agLNAs. Biomedical Relevance. There are currently few options for sequence-specific recognition of chromosomal DNA. agLNAs unlock access to chromosomal DNA and provide an alternative for addressing the difficult problem of achieving adequate potency and specificity in vivo. agLNAs will also provide useful tools for probing chromosome accessibility at specific sequences, information that may lead to a better understanding of replication, DNA repair, and gene expression. Chromosomal DNA encodes the information necessary to express proteins. Agents that target DNA have the potential to affect production of proteins involved in disease and may provide a new class of drugs. The goals of this proposal are to learn how recognition of DNA can be achieved and to use this knowledge to develop strategies for controlling the function of cells
Keywords: Achievement; Achievement Attainment; Address; Affect; Androgen Receptor; Anti-Sense Oligonucleotides; Antisense Agent; Antisense Oligonucleotides; Apo-B; ApoB; Apolipoproteins B; B-Cell CLL; B-Cell Chronic Lymphocytic Leukemia; B-Cell Chronic Lymphogenous Leukemia; B-Cell Chronic Lymphoid Leukemia; B-Lymphocytic Leukemia; B-Lymphocytic Leukemia, Chronic; C element; Carbon; Cell Function; Cell Process; Cell physiology; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cellular biology; Chemical Structure; Chromosomes; Chronic B-Cell Leukemias; Chronic Lymphatic Leukemia; Chronic Lymphocytic Leukemia; Chronic Lymphogenous Leukemia; Clinical; Clinical Treatment; Clinical Trials; Clinical Trials, Unspecified; Collaborations; Complementary DNA; Complex; Confocal Microscopy; D-Ribose; DNA; DNA Repair Gene; DNA, Complementary; Deoxyribonucleic Acid; Development; Digestion; Disease; Disorder; Drugs; Environment; Funding; Future; Gene Action Regulation; Gene Expression; Gene Expression Inhibitor; Gene Expression Regulation; Gene Organization; Gene Products, RNA; Gene Regulation; Gene Regulation Process; Gene Structure; Gene Structure/Organization; Gene Targeting; Genes; Goals; Human; Human, General; Investigators; Knowledge; LNA (nucleic acid); Label; Laboratories; Lead; Learning; Letters; Leukemia, B-Cell; Life; Lymphoblastic Leukemia, Chronic; Lymphocytic Leukemia, B-Cell; Lymphocytic Leukemia, Chronic, B-Cell; Man (Taxonomy); Man, Modern; Measures; Mediating; Medication; Messenger RNA; Methods; Microscopy, Confocal; Modeling; Modification; Molecular Target; Nucleic Acids; O element; O2 element; Oligo; Oligonucleotides; Oligonucleotides, Antisense; Oxygen; PNA; Pb element; Peptide Nucleic Acids; Peptides; Pharmaceutic Preparations; Pharmaceutical Agent; Pharmaceutical Preparations; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Production; Progesterone Receptors; Promoter; Promoter Regions; Promoter Regions (Genetics); Promoters (Genetics); Promotor; Promotor (Genetics); Promotor Regions; Promotor Regions (Genetics); Proteins; Protocol; Protocols documentation; RNA; RNA Sequences; RNA, Messenger; RNA, Non-Polyadenylated; Receptors, Progesterone; Receptors, Progestin; Recruitment Activity; Research; Research Design; Research Personnel; Researchers; Resistance; Ribonucleic Acid; Ribose; Sequences, RNA; Specificity; Structure-Activity Relationship; Study Type; Subcellular Process; Targetings, Gene; Temperature; Testing; Therapeutic; Transcript; antigene; base; cDNA; carbene; cell biology; chemical structure function; chronic lymphoid leukemia; clinical investigation; cost; design; designing; disease/disorder; drug/agent; experience; experiment; experimental research; experimental study; fluorophore; gene product; genetic promoter element; heavy metal Pb; heavy metal lead; improved; in vivo; insight; locked nucleic acid; mRNA; melting; methylene; methylene radical; new approaches; novel; novel approaches; novel strategies; novel strategy; nuclease; progesterone receptor; public health relevance; recruit; research study; resistant; structure function relationship; study design; tool; transcription factor; trial regimen; trial treatment; uptake
Relevance: Chromosomal DNA encodes the information necessary to express proteins. Agents that target DNA have the potential to affect production of proteins involved in disease and may provide a new class of drugs. The goals of this proposal are to learn how recognition of DNA can be achieved and to use this knowledge to develop strategies for controlling the function of cells
Project start date: 2005-02-01
Project end date: 2013-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-07-070
5R01GM073042-06 (2010): $298426
Recognition Of Chromosomal DNA By Peptide Nucleic Acids
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas Dallas, Tx 753909105
Grant 5R01GM073042-03 from National Institute Of General Medical Sciences IRG: ZRG1
Abstract: Peptide nucleic acid (PNA) is a DNA mimic that recognizes complementary sequences by Watson-Crick base-pairing. One of the strengths of PNAs is their ability to recognize sites within duplex DNA by strand invasion. We have developed efficient methods for synthesizing PNAs and introducing them into cells. We have also characterized strategies for improving the efficiency of strand invasion. We now propose to combine these advances to develop antigene PNAs as agents for manipulating gene expression at the level of the chromosome. The objective of this proposal is to understand the properties of antigene PNAs and test the hypothesis that antigene PNAs can recognize chromosomal DNA inside cells. Specifically, we propose to 1) Characterize and optimize the intracellular localization of PNAs, 2) Develop rules for using antisense PNAs to manipulate gene expression, and 3) Explore the value of antigene PNAs as a general strategy for recognizing duplex DNA by targeting genes that are important in the progression of cancer, specifically c-myc and the reverse transcriptase component of telomerase h TER T. To achieve these goals my laboratory will take advantage of our ability to rapidly synthesize PNAs and PNA-peptide conjugates and our experience with using PNAs inside cells. Preliminary experiments have already shown that PNAs can act as antigene agents, providing a powerful starting point for the experiments described in this proposal. Compounds that sequence-specifically recognize chromosomal DNA have enormous potential to treat disease and be powerful research tools. Such agents might block gene expression, activate gene expression, or enable harmful mutations to be corrected. In spite of the attractiveness of chromosomal DNA as a target, the development of antigene agents has been slow. The studies described in this proposal rigorously test the value of antigene PNAs for the recognition of chromosomes and our data will shape decisions regarding the use of PNAs for laboratory studies and clinical development.
Keywords: DNA, chromosome, nucleic acid quantitation /detection, peptide nucleic acid, gene expression, genetic manipulation, intracellular transport, oncogene, progesterone receptor, protooncogene, telomerase, confocal scanning microscopy, neoplastic cell, nucleic acid chemical synthesis
Project start date: 2005-02-01
Project end date: 2009-01-31
5R01GM073042-03 (2007): $244063
5R01GM073042-02 (2006): $251352
Grants awarded to David R Corey
RECOGNITION OF CHROMOSOMAL DNA BY DOUBLE-STRANDED RNA
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas, Dallas, Tx 75390-9105
Grant 3R01GM077253-03S1 from National Institute Of General Medical Sciences
Abstract: This competitive revision is being submitted for Notice Number (NOT-OD-09-058) and Notice Title NIH Announces the Availablity of Recovery Act Funds for Competitive Revision Applications. Background. The ability of short interfering RNAs (siRNAs) to recognize mRNA is widely appreciated. siRNAs are entering Phase 1 trials, but their ultimate potential to impact human health is unclear. The overall goal of the parent proposal GM 77253 was to investigate the action of duplex RNAs that are complementary to gene promoters. GM 77253 had four Aims i. Evaluate involvement of argonaute proteins in RNA-mediated gene activation ii. Characterize the mechanism of transcriptional silencing by agRNAs iii. Define rules for targeting gene promoters iv. Identify endogenous miRNAs that inhibit transcription Rationale for a significant expansion of research scope. Recent transcriptome studies have revealed many noncoding transcripts overlapping the 3´ terminus of genes. The function of these transcripts is unknown and the potential for regulation downstream from the 3´-UTR has attracted little attention. While pursuing Aim ii, we observed that promoter-targeted RNAs were binding to a noncoding transcript that overlaps the 5´ terminus of our target gene progesterone receptor. We hypothesized that RNAs that target sequences past the 3´-terminus of genes might also be able to bind noncoding transcripts and modulate gene expression. We characterized transcription at the progesterone receptor (PR) locus and identified noncoding transcripts that overlap the 3´ end of the gene. Small RNAs complementary to a noncoding transcript inhibit PR transcription in T47D cells (a cell line with high PR expression) and activate PR transcription in MCF7 cells (a cell line with low PR expression). The RNAs recruit argonaute to the 3´- noncoding transcript, alter levels of RNA polymerase, and modulate transcription of PR pre-mRNA and mature mRNA. RNAs complementary to sequences beyond the 3´ mRNA terminus of BRCA1 also inhibit gene expression. Our results extend the potential for RNA-mediated gene regulation to regions downstream from the 3´ end of genes. We propose to characterize the mechanism of action of 3´-targeted RNAs to understand how they can modulate transcription even though they are not complementary to mRNA and target sequences that are far distant from gene promoters. Regulation of gene expression is fundamental to biological processes. Our research investigates regions of chromosomal DNA that lie beyond the termini of genes. These regions have received little attention and we will investigate whether gene regulation can extend to these regions
Keywords: Activation, Gene; Affect; Androgen Receptor; Attention; BRCA1; BRCA1 gene; Binding; Binding (Molecular Function); Biological Function; Biological Process; Breast Cancer 1 Gene; Breast Cancer 1, Early Onset Gene; Breast Cancer Type 1 Susceptibility Gene; CRNA; Cell Line; Cell Lines, Strains; CellLine; Cells; Chromosomes; Clinical Trials, Phase I; Complementary RNA; DNA; DNA-Dependent RNA Polymerases; DNA-Directed RNA Polymerase; Data; Deoxyribonucleic Acid; Distant; Double-Stranded RNA; EC 2.7.7.6; Early-Stage Clinical Trials; Figs; Figs - dietary; Funding; Genbank; Gene Action Regulation; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Inactivation; Gene Products, RNA; Gene Regulation; Gene Regulation Process; Gene Silencing; Gene Targeting; Gene Transcription; Genes; Genetic Transcription; Goals; Health; Hereditary Breast Cancer 1; Human; Human, General; Immune Precipitation; Immunoprecipitation; Laboratories; Learning; Liver; MCF-7; MCF-7 Cell; MCF7; MCF7 cell; Man (Taxonomy); Man, Modern; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Messenger RNA; Molecular Interaction; NIH; National Institutes of Health; National Institutes of Health (U.S.); Nucleoside-triphosphate[{..}]RNA nucleotidyltransferase (DNA-directed); PSCP; Parents; Phase 1 Clinical Trials; Phase I Clinical Trials; Phase I Study; Post-Transcriptional Gene Silencing; Post-Transcriptional Gene Silencings; Posttranscriptional Gene Silencing; Posttranscriptional Gene Silencings; Pre-mRNA; Preparation; Progesterone Receptors; Promoter; Promoter Regions; Promoter Regions (Genetics); Promoters (Genetics); Promotor; Promotor (Genetics); Promotor Regions; Promotor Regions (Genetics); Proteins; Quelling; RNA; RNA Expression; RNA Interference; RNA Polymerases; RNA Silencing; RNA Silencings; RNA, Double-Stranded; RNA, Messenger; RNA, Messenger, Precursors; RNA, Non-Polyadenylated; RNA, Noncoding; RNA, Nontranslated; RNA, Untranslated; RNAi; RNF53; Receptor Protein; Receptors, Progesterone; Receptors, Progestin; Recovery; Recruitment Activity; Regulation; Research; Ribonucleic Acid; Sequence-Specific Posttranscriptional Gene Silencing; Small RNA; T47D; Targetings, Gene; Terminator Regions; Terminator Regions (Genetics); Terminator Sequence; Testing; Therapeutic Agents; Transcript; Transcription; Transcription, Genetic; Transcriptional Terminator Regions; UTRs; United States National Institutes of Health; Untranslated RNA; Untranslated Regions; Work; base; body system, hepatic; brca 1 gene; cultured cell line; dsRNA; gene expression signature; gene product; genetic promoter element; genetic terminator element; mRNA; mRNA Precursor; organ system, hepatic; phase 1 study; phase 1 trial; phase I trial; premRNA; progesterone receptor; protocol, phase I; public health relevance; receptor; receptor expression; recruit; tool; transcriptome
Relevance: Narrative Regulation of gene expression is fundamental to biological processes. Our research investigates regions of chromosomal DNA that lie beyond the termini of genes. These regions have received little attention and we will investigate whether gene regulation can extend to these regions
Project start date: 2009-09-30
Project end date: 2011-07-31
Budget start date: 30-SEP-2009
Budget end date: 31-JUL-2011
PFA/PA: PA-07-070
3R01GM077253-03S1 (2009): $341714
5R01GM077253-04 (2010): $295317
5R01GM077253-03 (2009): $298300
5R01GM077253-02 (2008): $298300
1R01GM077253-01A2 (2007): $298300
NEW CHEMISTRIES TO INHIBIT TELOMERASE IN CANCER CELLS
David R Corey, Professor
Pharmacologyuniversity Of Texas Sw Med Ctr/dallas
dallas, Tx 753909105
Grant 5R01CA074908-03 from National Cancer Institute IRG: CTY
Abstract: Telomerase activity has been found in 85-95 percent of all primary tumors that have been assayed, but not in somatic cells with the exception of germ cells and proliferative cells of renewal tissues (which express low levels). The connection between telomerase and cancer has led to the hypothesis that telomerase activity is necessary for sustained proliferation of most cancer cells, and that telomerase inhibitors might be effective chemotherapeutic agents. The objective of this proposal is the development of telomerase inhibitors as tools to validate the connection between telomerase and cancer. Such inhibitors would be lead compounds for the design of anti-telomerase therapeutics and further the understanding of the interplay between telomerase and other cellular proteins. Specific aims of this proposal include the discovery and optimization of new inhibitor chemistries in vitro, the use of inhibitors as structural and functional probes for telomerase, and the use of the inhibition of telomerase activity to help resolve whether telomerase activity is necessary for cell immortality
Keywords: chemical kinetics, enzyme inhibitor, nucleic acid chemical synthesis, peptide chemical synthesis, telomerase DNA, RNA, cell growth regulation, oligonucleotide
Project start date: 1998-04-01
Project end date: 2001-01-31
5R01CA074908-03 (2000): $81310
5R01CA074908-02 (1999): $78942
Recognition Of Chromosomal DNA By Chemically Modified Oligonucleotides
David R Corey, Professor
Pharmacologyuniversity Of Texas Sw Med Ctr/dallas
Grant 2R01GM073042-05 from National Institute Of General Medical Sciences IRG: ZRG1
Abstract: Synthetic agents that recognize specific sequences within chromosomal DNA are a promising strategy for controlling gene expression. During the previous funding period we described duplex RNAs, peptide nucleic acids (PNAs), PNA-peptides, and locked nucleic acids (LNAs) that target gene promoters and modulate expression. LNA bases contain a modified ribose with a methylene linkage between the 2´-oxygen and 4´-carbon and increase melting temperature values (Tm´s) by as much as 100C per substitution. We focus this proposal on LNAs because LNAs have been the simplest and most effective tools for direct recognition of chromosomal DNA. Objectives. Our objective for the next funding period is to understand the potential of chromosome- targeted LNAs as research tools and leads for clinical development. Research Design. We will investigate the mechanism, optimization and applications of agLNAs. In Aim 1 we will investigate the mechanism of agLNA-mediated recognition of nucleic acid targets and subsequent inhibition of gene expression inside cells. The environment surrounding gene promoters is a complex mix of RNA transcripts, proteins, and chromosomal DNA. Understanding the mechanism of recognition will help guide use of agLNAs for new applications and provide important basic insights into how synthetic oligomers affect cellular processes at gene promoters. In Aim 2, we will characterize cellular uptake, improve delivery methods, and test chemically modified LNAs. The goal for this Aim is to identify the most straightforward and efficient combination of delivery protocol and chemical structure for achieving optimal inhibition with antigene oligonucleotides. This information will assist researchers hoping to use agLNAs and help guide the design of agLNAs for therapeutic applications. In Aim 3 we propose novel strategies to use agLNAs to control cellular processes. Applications include use of agLNAs to probe accessibility of sequences within chromosomal DNA, targeting agLNAs to genes with bidirectional promoters (10 % of human genes), using agLNAs to manipulate expression of genes that are important biomedical targets, and synthesis and testing of agLNA-peptide conjugates designed to recruit transcription factors to gene promoters and activate gene expression. These experiments were chosen for their potential to expand the range of applications for agLNAs. Biomedical Relevance. There are currently few options for sequence-specific recognition of chromosomal DNA. agLNAs unlock access to chromosomal DNA and provide an alternative for addressing the difficult problem of achieving adequate potency and specificity in vivo. agLNAs will also provide useful tools for probing chromosome accessibility at specific sequences, information that may lead to a better understanding of replication, DNA repair, and gene expression. Chromosomal DNA encodes the information necessary to express proteins. Agents that target DNA have the potential to affect production of proteins involved in disease and may provide a new class of drugs. The goals of this proposal are to learn how recognition of DNA can be achieved and to use this knowledge to develop strategies for controlling the function of cells
Project start date: 2005-02-01
Project end date: 2013-01-31
Controlling Gene Expression With Peptide Nucleic Acids
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas Dallas, Tx 753909105
Grant 5R01GM060642-08 from National Institute Of General Medical Sciences IRG: ZRG1
Abstract: Peptide nucleic acid (PNA) is a DNA/RNA mimic that recognizes complementary sequences by Watson-Crick base-pairing and binds to RNA with high affinity. We have developed efficient methods for synthesizing PNAs and introducing them into cells. Once inside cells, we have shown that PNAs can efficiently inhibit gene expression. The long-term objective of this proposal is to better understand the properties of antisense PNAs and test the hypothesis that PNAs can be useful agents for probing biological function inside cells. Specifically, we propose to 1) Characterize the rules governing successful use of antisense PNAs to inhibit gene expression, 2) Use antisense PNAs to examine the consequences of inhibiting significant biological targets, thereby demonstrating that PNAs can be useful tools for studying cellular phenotypes, and 3) Use PNAs to detect gene expression inside cells. To achieve these goals my laboratory will take advantage of our ability to rapidly synthesize PNAs and PNA-peptide conjugates and our experience with using PNAs inside cells. The potential for compounds that recognize nucleic acids to treat disease and be powerful research tools is enormous. Few, if any, other approaches to drug discovery can move through the initial stages of development so rapidly. In spite of compelling advantages, progress in the clinic has been slow. Antisense PNAs possess dramatically different chemical properties relative to RNA or DNA oligomers that can also block gene expression. These chemical differences may confer advantages upon PNA and encourage the development of new research tools or more effective therapeutics. The work proposed here will rigorously test the value of antisense PNAs and our data will shape decisions regarding the use of PNAs for laboratory studies and clinical development.
Keywords: antisense nucleic acid, gene expression, nucleic acid hybridization, nucleotide analog, oligonucleotide, peptide analog, Coronaviridae, RNA splicing, biomimetics, cell biology, gene induction /repression, hepatitis C virus, messenger RNA, molecular probe, nonmammalian vertebrate embryology, phenotype, progesterone receptor, virus infection mechanism, virus replication, zebrafish, RNA interference
Project start date: 2000-07-01
Project end date: 2008-06-30
5R01GM060642-08 (2007): $266250
5R01GM060642-07 (2006): $326208
Sponsored Links Excellgen http://Excellgen.com
5R01GM060642-06 (2005): $334058
2R01GM060642-05 (2004): $280800
Recognition Of Chromosomal DNA By Peptide Nucleic Acids
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas Dallas, Tx 753909105
Grant 1R01GM073042-01 from National Institute Of General Medical Sciences IRG: ZRG1
Abstract: Peptide nucleic acid (PNA) is a DNA mimic that recognizes complementary sequences by Watson-Crick base-pairing. One of the strengths of PNAs is their ability to recognize sites within duplex DNA by strand invasion. We have developed efficient methods for synthesizing PNAs and introducing them into cells. We have also characterized strategies for improving the efficiency of strand invasion. We now propose to combine these advances to develop antigene PNAs as agents for manipulating gene expression at the level of the chromosome. The objective of this proposal is to understand the properties of antigene PNAs and test the hypothesis that antigene PNAs can recognize chromosomal DNA inside cells. Specifically, we propose to 1) Characterize and optimize the intracellular localization of PNAs, 2) Develop rules for using antisense PNAs to manipulate gene expression, and 3) Explore the value of antigene PNAs as a general strategy for recognizing duplex DNA by targeting genes that are important in the progression of cancer, specifically c-myc and the reverse transcriptase component of telomerase h TER T. To achieve these goals my laboratory will take advantage of our ability to rapidly synthesize PNAs and PNA-peptide conjugates and our experience with using PNAs inside cells. Preliminary experiments have already shown that PNAs can act as antigene agents, providing a powerful starting point for the experiments described in this proposal. Compounds that sequence-specifically recognize chromosomal DNA have enormous potential to treat disease and be powerful research tools. Such agents might block gene expression, activate gene expression, or enable harmful mutations to be corrected. In spite of the attractiveness of chromosomal DNA as a target, the development of antigene agents has been slow. The studies described in this proposal rigorously test the value of antigene PNAs for the recognition of chromosomes and our data will shape decisions regarding the use of PNAs for laboratory studies and clinical development.
Keywords: DNA, chromosome, nucleic acid quantitation /detection, peptide nucleic acid, gene expression, genetic manipulation, intracellular transport, oncogene, progesterone receptor, protooncogene, telomerase, confocal scanning microscopy, neoplastic cell, nucleic acid chemical synthesis
Project start date: 2005-02-01
Project end date: 2009-01-31
1R01GM073042-01 (2005): $257400
CONTROLLING GENE EXPRESSION WITH PEPTIDE NUCLEIC ACIDS
David R Corey, Professor
Pharmacologyuniversity Of Texas Sw Med Ctr/dallas
dallas, Tx 753909105
Grant 5R01GM060642-02 from National Institute Of General Medical Sciences IRG: MEDB
Abstract: Genome sequencing is expanding our knowledge of the sequence of cellular proteins, DNA, and RNA at a pace that surpasses our understanding of their function. Sequence data alone cannot elucidate the role of a protein in cellular processes and pathways, but can be used to design an oligonucleotide-based antisense or antigene approach as a simple route to "knock out" phenotypes. Many laboratories have tried this, but their success has been limited because it has been difficult to predictably design oligonucleotides that function in a potent and selective manner. Peptide nucleic acids (PNAs) are oligonucleotide mimics that possess a nonionic amide backbone. They have been demonstrated to possess substantial advantages for hybridization in cell free systems, and the goal of this proposal is to determine whether they possess similar advantages for hybridization within cells. The investigators specific aims are to (i) optimize delivery of PNAs into cells, (ii) use PNAs to control gene expression by targeting mRNA, and (iii) use PNAs to control gene expression by targeting genomic DNA. Their research is relevant to human health because the substantial advantages that PNAs possess for hybridization to complementary targets suggest that they will be useful tools for translating genome data into knowledge of the details of protein function within cells. In addition, PNAs may provide a new generation of antisense agents that improve on first generation therapeutics that have already shown promise in clinical trials. Information generated by this proposal will facilitate evaluation of PNAs as a strategy for targeting human disease and provide a powerful approach for translating the one dimensional understanding of protein function derived from genomic data into the multidimensional understanding necessary to understand cell signaling and regulation
Keywords: antisense nucleic acid, gene expression, nucleic acid hybridization, nucleotide analog, oligonucleotide, peptide analog
Project start date: 2000-07-01
Project end date: 2004-06-30
5R01GM060642-02 (2001): $234733
1R01GM060642-01 (2000): $229637
CONSEQUENCES OF INHIBITION OF CELLULAR TELOMERASE
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas Dallas, Tx 753909105
Grant 5R01CA085363-04 from National Cancer Institute IRG: CDF
Abstract: Adapted from s ) Telomerase activity has been found in most types of human tumors, but not in adjacent normal cells. This correlation has led to the hypothesis that reactivation of telomerase is necessary for the sustained proliferation that characterizes cancer, and that telomerase is a novel target for chemotherapy. The validity of this hypothesis has been vigorously debated, and effective inhibitors of telomerase are needed to understand how blocking telomerase activity will affect cancer cell phenotypes and proliferation. To discover the potential for telomerase as a target for chemotherapy Dr. Corey is developing oligonucleotides and oligonucleotide mimics as highly selective and potent inhibitors. These oligomers are well suited to achieving unambiguous insights into the consequences of telomerase inhibition because they take advantage of the potential for stringently selective recognition inherent in Watson-Crick base-pairing. An oligomer complementary to a target sequence should exert a sequence-specific physiologic effect, whereas the mismatch containing oligomer should not. They now propose to continue to develop synthetic telomerase inhibitors, characterize the effects of telomerase inhibition on the proliferation of varied lines of cultured cells, and perform preclinical studies in mouse models of human cancer. This research will be integrated into a cycle of experiments in which the observed effects of inhibitors in cell culture and animals will rapidly lead to revised inhibitor designs and renewed testing for efficacy. These studies will characterize the in vivo effects of inhibitors of telomerase, permit investigation of the side effects of antitelomerase therapy, and provide insights that will be generally applicable to the development of all classes of telomerase inhibitors.
Keywords: breast neoplasm, enzyme inhibitor, lung neoplasm, neoplasm /cancer chemotherapy, nonhuman therapy evaluation, prostate neoplasm, telomerase, apoptosis, cell proliferation, drug design /synthesis /production, enzyme activity, oligonucleotide, SCID mouse, athymic mouse, fluorescent in situ hybridization
Project start date: 2001-02-08
Project end date: 2005-01-31
5R01CA085363-04 (2004): $351000
1R01CA085363-01A1 (2001): $351000
Controlling Gene Expression With Peptide Nucleic Acids
David R Corey, Professor
University Of Texas Sw Med Ctr/dallas Dallas, Tx 753909105
Grant 3R01GM060642-05S1 from National Institute Of General Medical Sciences IRG: ZRG1
Project start date: 2000-07-01
Project end date: 2008-06-30
3R01GM060642-05S1 (2004): $53258