Lorem Sit Amet Dolor
Researcher: Lorem Sit Amet

123 abc lane, Townsville, ZZ 00000, USA
Funding Progress: $§ / $§§§§§

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Controlling DIPG/DMG with the protein kinase C-iota inhibitor ICA-1S
Translational
DIPG, Childhood (Brain Cancer)
Lay Summary

Pediatric Diffuse Intrinsic Pontine Glioma (DIPG) and Diffuse Midline Glioma (DMG) are currently incurable childhood brain tumors. Elucidating the regulation of signaling pathways in pediatric DIPG and DMG is critical to targeting tumor cell migration and spreading.  Until these pathways are understood, comprehensive and effective treatment of this group of incurable primary brain tumors may remain elusive.  This research focuses on Protein Kinase C-iota (PKC-i) which plays a major role in tumorigenesis because it is overexpressed and impacted by phosphoinositide 3-kinase (PI3K), phosphatase and tensin homolog (PTEN) and other signaling pathways.  However, there is a gap in knowledge regarding which specific PKC isozymes are responsible for pediatric DIPG and DMG cell migration and which cell signaling pathways are regulated by PKC-i in pediatric DIPG and DMG. We have identified a novel PKC-i inhibitor ICA-1S [1] that inhibits PKC-i activity and proliferation of adult Glioblastoma Multiforme (GBM) in-vitro and in-vivo in mice tumor models [2].  An objective here is to establish whether ICA-1S alone or in combination with temozolomide (TMZ) decreases in-vitro pediatric DIPG and DMG cell migration by blocking the PKC-i induced regulation of focal adhesion kinase (FAK) and paxillin [2].  The central hypothesis is that a PKC-i inhibitor (ICA-1S) alone or in combination with TMZ can restrain pediatric DIPG and DMG cell migration by intercepting the PKC-i/FAK/paxillin cascade in-vitro. We intend to test our central hypothesis and, thereby achieve the objective of this proposal by addressing the following two specific aims: 1) Establish whether PKC-i protein/or mRNA is over-expressed in pediatric DIPG and DMG biopsies compared to normal pediatric brain tissue; 2) Determine the in-vitro effects of ICA-1S alone and in combination with TMZ on cell migration, apoptosis, PKC activity, PKC-i/FAK/paxillin pathway in pediatric DIPG and DMG cell lines. Hence, the proposed research is innovative because it will identify an inhibitor for PKC-i that inhibits pediatric DIPG and DMG cell migration and cell spreading.  As a consequence, we envision that this research may advance the ability to treat DIPG and DMG, because the amount of PKC-i in DIPG and DMG specimens could be used as a biomarker for children who may benefit from anti-PKC-i therapy with ICA-1S or one of its derivatives.  This accomplishment would be a significant advancement for pediatric personalized medicine. 

Executive Summary

1. Pediatric Diffuse Intrinsic Pontine Glioma (DIPG) and Diffuse Midline Glioma (DMG) are currently incurable childhood brain tumors. Elucidating the regulation of signaling pathways in pediatric DIPG and DMG is critical to targeting tumor cell migration and spreading.  Until these pathways are understood, comprehensive and effective treatment of this group of incurable primary brain tumors may remain elusive.  This research focuses on Protein Kinase C-iota (PKC-i) which plays a major role in tumorigenesis because it is overexpressed and impacted by phosphoinositide 3-kinase (PI3K), phosphatase and tensin homolog (PTEN) and other signaling pathways.  However, there is a gap in knowledge regarding which specific PKC isozymes are responsible for pediatric DIPG and DMG cell migration and which cell signaling pathways are regulated by PKC-i in pediatric DIPG and DMG. We have identified a novel PKC-i inhibitor ICA-1S [1] that inhibits PKC-i activity and proliferation of adult Glioblastoma Multiforme (GBM) in-vitro and in-vivo in mice tumor models [2].  An objective here is to establish whether ICA-1S alone or in combination with temozolomide (TMZ) decreases in-vitro pediatric DIPG and DMG cell migration by blocking the PKC-i induced regulation of focal adhesion kinase (FAK) and paxillin [2].  The central hypothesis is that a PKC-i inhibitor (ICA-1S) alone or in combination with TMZ can restrain pediatric DIPG and DMG cell migration by intercepting the PKC-i/FAK/paxillin cascade in-vitro. We intend to test our central hypothesis and, thereby achieve the objective of this proposal by addressing the following two specific aims: 1) Establish whether PKC-i protein/or mRNA is over-expressed in pediatric DIPG and DMG biopsies compared to normal pediatric brain tissue; 2) Determine the in-vitro effects of ICA-1S alone and in combination with TMZ on cell migration, apoptosis, PKC activity, PKC-i/FAK/paxillin pathway in pediatric DIPG and DMG cell lines.  The experimental methods that will be use to determine whether ICA-1can restrain DIPG/DMG proliferation and migration are: 1) cell biology techniques including cell cycle analysis using propidium iodine, bromodeoxyuridine and flow cytometry, tumor growth measurements, cell migration assays and immunofluorescent microscopy;  2) biochemical procedures including cell fractionation, PKC activity assays, immunoprecipitations, and Western blotting. Hence, the proposed research is innovative because it will identify an inhibitor for PKC-i that inhibits pediatric DIPG and DMG cell migration and cell spreading.  As a consequence, we envision that this research may advance the ability to treat DIPG and DMG, because the amount of PKC-i in DIPG and DMG specimens could be used as a biomarker for children who may benefit from anti-PKC-i therapy with ICA-1S or one of its derivatives.  This accomplishment would be a significant advancement for pediatric personalized medicine.

2.  We have published a similar study utlizing glioblastoma demonstrating the feasibilty of performing this investigation.

3. The goal of the proposed research is to investigate whether a novel PKC-i inhibitor (ICA-1S) identified in my laboratory can restrain DIPG and DMG spreading by intercepting the PKC-i/FAK/paxillin pathway in-vitro.  I (Mildred Acevedo-Duncan) have the expertise, leadership, and motivation necessary to successfully carry out the proposed research since I have studied protein kinase C (PKC) for more than 30 years.  My initial introduction to PKC was in 1987 in the laboratory of my post-doctoral mentor Robert Farese, MD. As a post-doctoral fellow at the VA Hospital in Tampa, I studied how PKCs were regulated and made antibodies to PKCs. I began independent research by investigating the role of PKC in carcinogenesis when I obtained my first grant from the H.L. Moffitt Cancer Center in 1992.  As a PI on two Veterans’ Administration Merit Review Grants, an industry grant (CRADA) and Foundation grants, I laid the groundwork for the proposed research by identifying PKC isozymes and PKC substrates which regulate the cell cycle to target and interfere with their activity, thereby identifying a selective therapeutic agent for controlling cancer cell proliferation/spreading/survival.  I am well suited as the PI for this project since I have discovered that PKC-i is overexpressed in a variety of cancers, have described the PKC-ι/Cdk7/cdk2, PKC-ι/IKKa/b/Ikba/NFkB, PKC-z/Rac1/Pak1/b-Catenin, PKC-i/FAK/paxillin pathway and PKC-ι/Bad pathways and have identify a novel PKC-ι inhibitor (ICA-1S).  I have published fourteen publications establishing the role of PKCs in glioblastoma proliferation, migration and survival (Cellular Signalling (2021) 77:109819, Neurochemical Research (2014) 39(9):1691-1701, Carcinogenesis. (2012) 33(1):10-19, Biochimica et Biophysica Acta Molecular Cell Research (2012)1813:1190–1197, Cell Proliferation (2008) 41:122-135, Cell Proliferation (2005)38:87-106. Tissue and Cell (2005) 37:53-58, Interna. Immunopharm. (2004) 4:1775-1784, Cell Proliferation (2002) 35:23-36, Cell Proliferation (2001) 34:31-41, Tissue & Cell (2001) 33:55-62, Neurochemical Research (1997) 22:775-984, Cell Growth and Differentiation (1995) 6: 1353-1365, Biochem. Biophys. Res. Commun. (1994) 205:127-134). I successfully administered the projects (e.g., staffing, research protections, budgeted), coordinated and collaborated with other researchers, collated the experimental data, interpreted results, wrote manuscripts and produced several peer-reviewed publications from each project.  My extensive expertise in microscopy, biochemistry, molecular biology and the methodologies used to analyze the cell proliferation/migration/survival and PKC indicate the strength of this program.  The current application builds logically on my prior work. In summary, my experience and expertise have prepared me to the lead the proposed project.

Description of Research Proposal

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Budget

Lorem ipsum dolor sit amet, consectetur adipiscing elit. Integer gravida non felis non euismod. Fusce finibus aliquet consequat. Nam ac metus bibendum, iaculis purus sed, suscipit ligula. Proin et nisi libero. Mauris non urna urna. Nullam augue eros, fringilla sed mauris vitae, porta tincidunt risus. Aliquam sed tincidunt sem. Quisque lacinia quam tortor, imperdiet efficitur odio iaculis in. Sed ultricies condimentum volutpat. Vivamus dignissim faucibus porta.

Curabitur ut ipsum non odio malesuada vulputate. Morbi maximus, est eu lobortis molestie, tortor sapien hendrerit nisi, in cursus odio diam ut odio. Fusce pulvinar volutpat velit. Aliquam erat volutpat. Integer rhoncus mollis suscipit. Praesent non ipsum mollis, finibus nunc a, scelerisque nibh. In feugiat iaculis velit, eu semper lacus dignissim nec. Praesent vitae nisi leo. Cras venenatis dictum magna ut semper. Sed eget eros nibh. Sed vitae quam sed dolor faucibus elementum. Curabitur interdum porttitor finibus. Nullam tincidunt odio lectus, sit amet rhoncus libero dapibus sed. Sed mollis egestas enim, vel porta tortor volutpat eget.

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Collaborations and Conflicts of Interest

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Curabitur ut ipsum non odio malesuada vulputate. Morbi maximus, est eu lobortis molestie, tortor sapien hendrerit nisi, in cursus odio diam ut odio. Fusce pulvinar volutpat velit. Aliquam erat volutpat. Integer rhoncus mollis suscipit. Praesent non ipsum mollis, finibus nunc a, scelerisque nibh. In feugiat iaculis velit, eu semper lacus dignissim nec. Praesent vitae nisi leo. Cras venenatis dictum magna ut semper. Sed eget eros nibh. Sed vitae quam sed dolor faucibus elementum. Curabitur interdum porttitor finibus. Nullam tincidunt odio lectus, sit amet rhoncus libero dapibus sed. Sed mollis egestas enim, vel porta tortor volutpat eget.