2007 |
Vancurova, Ivana |
R15Activity Code Description: Supports small-scale research projects at educational institutions that provide baccalaureate or advanced degrees for a significant number of the Nation’s research scientists but that have not been major recipients of NIH support. The goals of the program are to (1) support meritorious research, (2) expose students to research, and (3) strengthen the research environment of the institution. Awards provide limited Direct Costs, plus applicable F&A costs, for periods not to exceed 36 months. This activity code uses multi-year funding authority; however, OER approval is NOT needed prior to an IC using this activity code. |
Nuclear Translocation of I-Kappab-Alpha as a Therapeutic Target
[unreadable] DESCRIPTION (provided by applicant): NFkB is a critical transcription factor regulating expression of pro-inflammatory and anti- apoptotic genes. Previous studies from our laboratory demonstrated that as opposed to other cell types, resting human neutrophils contain predominant amount of NFkB inhibitor, IkBa, in the nucleus, and this increased nuclear accumulation of IkBa results in the inhibition of NFkB activity and increased rate of neutrophil apoptosis. Our recent data have shown that proteasome inhibition induces translocation of IkBa to the nucleus in leukemia HL-60 and U- 937 cells, and in cancer HeLa cells; however, the mechanisms are currently unknown. The central hypothesis of this proposal is that induction of nuclear accumulation of IkBa inhibits NFkB activity and expression of NFkB-regulated anti-apoptotic and pro-inflammatory genes, and could thus provide a basis for novel anti-cancer and anti-inflammatory therapies aimed at the inhibition of NFkB activity by the nuclear IkBa. The specific aims focus on analyzing the mechanisms by which the nuclear IkBa inhibits NFkB activity and expression of NFkB-regulated genes in leukemia HL-60 and U-937 cells. In Aim 1, we will test the hypothesis that the proteasome inhibition-induced nuclear translocation of IkBa results in the inhibition of NFkB activity and decreased expression of NFkB-regulated anti-apoptotic genes, by using transfection of HL-60 and U-937 cells with inhibitory IkBa RNA. In Aim 2, we will use chromatin immunoprecipitation to test the hypothesis that the nuclear IkBa inhibits NFkB activity by associating with the promoters of NFkB-regulated genes. In Aim 3, we will investigate the mechanisms that regulate the proteasome inhibition-induced nuclear translocation of IkBa. Identification of the key molecular events that control NFkB activity by the nuclear IkBa will broaden our understanding of the mechanisms regulating NFkB activity, and might provide a new class of drug targets to regulate the NFkB driven pro-inflammatory and anti-apoptotic gene expression. The focus of this proposal is to identify the mechanisms by which the nuclear translocation of IkBa regulates activity of the transcription factor NFkB. Since NFkB activity is increased in many human diseases including inflammatory disorders, cancer, and leukemia, identification of the mechanisms by which the nuclear IkBa inhibits NFkB activity will contribute to the development of novel anti-cancer and anti-inflammatory therapies. [unreadable] [unreadable] [unreadable]
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1 |
2010 |
Vancurova, Ivana |
R15Activity Code Description: Supports small-scale research projects at educational institutions that provide baccalaureate or advanced degrees for a significant number of the Nation’s research scientists but that have not been major recipients of NIH support. The goals of the program are to (1) support meritorious research, (2) expose students to research, and (3) strengthen the research environment of the institution. Awards provide limited Direct Costs, plus applicable F&A costs, for periods not to exceed 36 months. This activity code uses multi-year funding authority; however, OER approval is NOT needed prior to an IC using this activity code. |
Gene Specific Regulation of Nfkb by Nuclear Ikba in Inflammation and Cancer
DESCRIPTION (provided by applicant): Our long-term goal is to understand the mechanisms regulating transcription of NF?B-dependent anti-apoptotic and pro-inflammatory genes. Since NF?B activity and expression of NF?B-regulated pro-inflammatory and anti-apoptotic genes are increased in many inflammatory diseases, as well as in many types of human cancer and leukemia, inhibition of NF?B-dependent transcription thus represents an important therapeutic target. However, one of the main concerns regarding the NF?B inhibitors is their specificity, since many steps leading to NF?B activation are important for other cellular functions as well. Our previous studies have shown that an induction of nuclear translocation and accumulation of the NF?B inhibitor, I?B?, inhibits the in vitro NF?B DNA binding activity and induces apoptosis in human leukocytes and cancer cells. Importantly, our recent data indicate that the in vivo inhibition of NF?B-dependent transcription by nuclear I?B? is gene specific: while the transcription of pro-inflammatory cytokines TNF?, IL-12 and IL-6 in human leukocytes is inhibited by nuclear I?B?, transcription of IL-8 is not. However, at present, the mechanisms that regulate the in vivo nuclear interaction of I?B? with NF?B proteins and NF?B- dependent promoters are unknown. The central hypothesis of this proposal is that the regulation of NF?B-dependent transcription of pro-inflammatory and anti-apoptotic genes by nuclear I?B? is gene specific, and could thus provide a basis for novel anti-inflammatory and anti-cancer therapies aimed at the specific inhibition of NF?B activity by nuclear I?B?. The specific aims focus on analyzing the mechanisms that regulate the in vivo binding of I?B? to NF?B dimers and NF?B promoters, thus regulating NF?B-dependent transcription. In Aim 1, we will measure by chromatin immunoprecipitation the recruitment of NF?B and I?B? proteins to NF?B-regulated promoters of pro-inflammatory and anti-apoptotic genes in human U937 macrophages and leukemia Hut-78 cells. We will determine whether the ability of nuclear I?B? to inhibit NF?B-dependent transcription depends on the subunit composition of NF?B dimers. In Aim 2, we will identify the mechanisms that regulate the nuclear I?B?-NF?B interaction in U937 macrophages and Hut-78 leukemia cells, and we will test the hypothesis that the responsible mechanisms involve post-translational modification(s) of p65 NF?B, and/or gene specific recruitment of the proteasome. Identification of the mechanisms that regulate the gene specific transcription of NF?B-dependent genes by nuclear I?B? might provide a new strategy that would use the nuclear I?B? as a specific regulator of NF?B-dependent transcription. This approach could be applicable in both anti-inflammatory and anti-cancer therapies. In addition, this project will enhance the research environment at St. John's University by providing undergraduate students with numerous opportunities to learn the fundamentals of biomedical research. PUBLIC HEALTH RELEVANCE: This proposal focuses on the mechanisms that regulate ability of the nuclear inhibitory protein I?B? to associate with NF?B-regulated promoters and inhibit transcription of NF?B-dependent inflammatory and pro-survival genes. Since NF?B activity is increased in many human diseases including inflammatory disorders, cancer, and leukemia, identification of the mechanisms by which the nuclear I?B? inhibits NF?B-dependent transcription will lead to the development of more specific anti-inflammatory and anti-cancer therapies.
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2013 |
Vancurova, Ivana |
R15Activity Code Description: Supports small-scale research projects at educational institutions that provide baccalaureate or advanced degrees for a significant number of the Nation’s research scientists but that have not been major recipients of NIH support. The goals of the program are to (1) support meritorious research, (2) expose students to research, and (3) strengthen the research environment of the institution. Awards provide limited Direct Costs, plus applicable F&A costs, for periods not to exceed 36 months. This activity code uses multi-year funding authority; however, OER approval is NOT needed prior to an IC using this activity code. |
Interleukin-8 Regulation by Proteasome and Nuclear Ikb in Cancer and Inflammation
DESCRIPTION (provided by applicant): Interleukin-8 (IL-8) is an inflammatory chemokine that has a crucial role in cancer progression through its induction of tumor cell proliferation, recruitment and activation of tumor-infiltrating leukocytes, angiogenesis, and metastasis. The expression of IL-8, as well as expression of many other inflammatory cytokines is regulated at the level of transcription by NF?B. However, studies from our laboratory indicate that the transcriptional regulation of IL-8 differs from the regulation of other NF?B-dependent genes. Specifically, we have found that the nuclear I?B? does not inhibit IL-8 expression in stimulated leukocytes, while it inhibits expression of other NF?B-dependent genes. In addition, our recent data show that the proteasome inhibition that is used as an anti-cancer therapy for its ability to inhibit expression of NF?B-dependent genes actually increases IL-8 expression in metastatic prostate and ovarian cancer cells and in human macrophages. However, the responsible mechanisms are largely unknown. The proposed research addresses the lack of knowledge on the regulation of IL-8 transcription by the proteasome inhibition and by nuclear I?B?. Our long-term goal is to understand the transcriptional mechanisms regulating expression of NF?B-responsive genes. The objective of this proposal is to determine how the proteasome and nuclear I?B? regulate transcription of IL-8, and how this regulation differs from other NF?B-dependent genes. The central hypothesis is that S536 p65 phosphorylation, specificity of the DNA sequence of the NF?B binding site, and/or the transcription factor EGR-1 render IL-8 unresponsive to the inhibition by I?B?, and increase IL-8 expression in response to the proteasome inhibition. Based on our preliminary data, the project will test two mutually non-exclusive models. In Aim 1, we will test the hypothesis that the IL-8 promoter is occupied predominantly by p65 homodimers phosphorylated on S536, and that the proteasome inhibition increases S536 p65 phosphorylation, resulting in the increased IL-8 transcription. In Aim 2, we will test the hypothesis that the IL-8 promoter sequence and EGR-1 involvement are responsible for the proteasome inhibition increased IL-8 expression and resistance of IL-8 transcription to I?B? inhibition. We will use metastatic prostate cancer PC-3 cells, ovarian cancer OVCAR-3 cells and stimulated U937 macrophages as models in both Aims. Since IL-8 promotes tumor cell growth, angiogenesis, metastasis, as well as activates leukocytes, understanding its regulation by proteasome and nuclear I?B? may have important clinical implications in cancers and inflammatory disorders characterized by excessive IL-8 expression. In addition, this project will enhance the research environment at St. John's University by providing motivated underprivileged students with numerous opportunities to learn the fundamentals of biomedical research.
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1 |
2016 |
Vancurova, Ivana |
R15Activity Code Description: Supports small-scale research projects at educational institutions that provide baccalaureate or advanced degrees for a significant number of the Nation’s research scientists but that have not been major recipients of NIH support. The goals of the program are to (1) support meritorious research, (2) expose students to research, and (3) strengthen the research environment of the institution. Awards provide limited Direct Costs, plus applicable F&A costs, for periods not to exceed 36 months. This activity code uses multi-year funding authority; however, OER approval is NOT needed prior to an IC using this activity code. |
Targeting Ikk and Hdac Mediated Il-8 Expression in Ovarian and Prostate Cancer
ABSTRACT Our long-term goal is to understand the transcriptional mechanisms that regulate the expression of NF?B- dependent chemokines as they relate to cancer development and progression. Interleukin-8 (IL-8, CXCL8) is a pro-inflammatory and pro-angiogenic chemokine that contributes to cancer progression through its induction of tumor cell survival, angiogenesis, and metastasis. IL-8 expression is increased in many types of advanced cancers, including ovarian (OC) and prostate (PC) cancer, and correlates with poor prognosis. There are no effective treatments for the advanced stages of ovarian and prostate cancer; thus, novel therapeutic strategies are urgently needed. Even though histone deacetylase (HDAC) inhibition by HDAC inhibitors (HDIs) and proteasome inhibition by bortezomib (BZ) have been highly effective in the treatment of cutaneous T cell lymphoma (CTCL) and other hematological cancers, they have been less effective in solid tumors, including ovarian and prostate cancer; the responsible mechanisms are not fully understood. Our preliminary studies show that while HDIs suppress the IL-8 expression in CTCL cells, they induce the IL-8 expression in solid tumors and monocytic cells. Furthermore, our results indicate that the HDI-induced IL-8 expression is partly mediated by I?B kinase (IKK), and that suppression of the HDI-induced IL-8 expression decreases survival of cancer cells. In addition, we have shown that while proteasome inhibition suppresses expression of most of the NF?B-dependent genes, it paradoxically increases the expression of IL-8 in cancer cells. Importantly, our recent in vivo data have demonstrated that combining BZ with anti- inflammatory IKK inhibitor significantly reduces ovarian tumor growth in mice when compared to either drug alone, indicating that using anti-inflammatory IKK inhibitors may increase BZ effectiveness in solid tumors. The central hypothesis, based on our preliminary data, is that anti-inflammatory therapy targeting the HDI- and BZ-induced IL-8 expression will increase HDI and BZ effectiveness in ovarian and prostate cancer. This hypothesis will be tested in two specific aims. In Aim 1, we will determine the in vitro mechanisms by which HDAC inhibition increases the IL-8 expression in ovarian and prostate cancer cells, and in monocytic cells. In Aim 2, we will test the hypothesis that anti-inflammatory therapy increases HDI and BZ effectiveness in reducing ovarian and prostate cancer growth in nude mice, and we will investigate the responsible in vivo mechanisms. Understanding the mechanisms by which HDAC and proteasome inhibition induces IL-8 expression will lead to the development of novel combination strategies for the treatment of ovarian and prostate cancer, and other drug resistant solid tumors characterized by excessive IL-8 release. In addition, this project will enhance the research environment at St. John's University by providing undergraduate and graduate students with numerous opportunities to learn the fundamentals of biomedical research.
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