{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/90460"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/90460","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Identification and characterization of a small molecule inhibitor of IMP-1 that decreases expression of IMP-1 target mRNAs and inhibits proliferation of IMP-1 positive cancer cells","abstract":"RNA-binding proteins control a variety of biological processes ranging from messenger RNA splicing to transport and translation. These post-transcriptional events are critical for proper cell function. One emerging class of proteins functions in several of these capacities. The VICKZ family of RNA-binding proteins is involved in translation control, mRNA localization and mRNA stability. I have studied the Insulin-like Growth Factor-2 mRNA-Binding Protein 1 (IGF2BP1/IMP-1/CRD-BP). IMP-1 exhibits an oncofetal pattern of expression, where it is expressed in embryonic development and its expression is repressed shortly after birth. However, the IMP-1 gene is reactivated in many different human cancers. Overexpression of IMP-1 leads to increased levels of proteins that promote tumor growth, metastasis, and resistance to anticancer drugs and is associated with a poor prognosis. IMP-1 enhances proliferation and migration of cancer cells by binding to and stabilizing mRNAs important in cancer, such as c-Myc. Although the role of c-Myc in cancer has been well established, it has remained an elusive therapeutic target because of its role as a transcription factor in non-neoplastic proliferating cells. Given its oncofetal pattern of expression, targeting IMP-1 presents a novel approach to targeting c-Myc. To identify new chemical entities with therapeutic potential in IMP-1 positive cancer, we carried out a pilot screen using an in vitro fluorescence anisotropy microplate assay (FAMA) and found that this approach was robust and appropriate for high throughput screening. We then carried out a high throughput screen of approximately 150,000 small molecules. Reported here is BTYNB, the first small molecule inhibitor of IMP-1, BTYNB decreases levels of IMP-1 target mRNAs, inhibits proliferation of IMP-1 positive cancer cells, and functions through the unique mechanism of decreasing oncogene mRNA stability. We believe that BTYNB not only can be developed as a potential therapeutic agent, but also serves as a useful molecular tool, with which we can probe the actions of IMP-1 in cancer cells. In addition to identifying and characterizing the first small molecule inhibitor of IMP-1, we were also interested in identifying novel molecular targets of IMP-1. Using in silico analysis of publicly available microarrays where IMP-1 was knocked down, we identified a panel of candidate target genes. Using qRTPCR and Western blot analysis, we then confirmed whether or not the mRNAs of candidate genes were decreased with IMP-1 knockdown and identified Protein Kinase C a (PKCa) as a new molecular target of IMP-1. Overall, this work has led to the identification and characterization of the first small molecule inhibitor of IMP-1 and has demonstrated that despite the fact that studies of the role of IMP-1 in cancer are rapidly expanding, there still remain novel molecular targets, such as PKCa, which may play critical roles in IMP-1 action in cancer cells.","abstract_html":"RNA-binding proteins control a variety of biological processes ranging from messenger RNA splicing to transport and translation. These post-transcriptional events are critical for proper cell function. One emerging class of proteins functions in several of these capacities. The VICKZ family of RNA-binding proteins is involved in translation control, mRNA localization and mRNA stability. I have studied the Insulin-like Growth Factor-2 mRNA-Binding Protein 1 (IGF2BP1/IMP-1/CRD-BP). IMP-1 exhibits an oncofetal pattern of expression, where it is expressed in embryonic development and its expression is repressed shortly after birth. However, the IMP-1 gene is reactivated in many different human cancers. Overexpression of IMP-1 leads to increased levels of proteins that promote tumor growth, metastasis, and resistance to anticancer drugs and is associated with a poor prognosis. IMP-1 enhances proliferation and migration of cancer cells by binding to and stabilizing mRNAs important in cancer, such as c-Myc. Although the role of c-Myc in cancer has been well established, it has remained an elusive therapeutic target because of its role as a transcription factor in non-neoplastic proliferating cells. Given its oncofetal pattern of expression, targeting IMP-1 presents a novel approach to targeting c-Myc. To identify new chemical entities with therapeutic potential in IMP-1 positive cancer, we carried out a pilot screen using an in vitro fluorescence anisotropy microplate assay (FAMA) and found that this approach was robust and appropriate for high throughput screening. We then carried out a high throughput screen of approximately 150,000 small molecules. Reported here is BTYNB, the first small molecule inhibitor of IMP-1, BTYNB decreases levels of IMP-1 target mRNAs, inhibits proliferation of IMP-1 positive cancer cells, and functions through the unique mechanism of decreasing oncogene mRNA stability. We believe that BTYNB not only can be developed as a potential therapeutic agent, but also serves as a useful molecular tool, with which we can probe the actions of IMP-1 in cancer cells. In addition to identifying and characterizing the first small molecule inhibitor of IMP-1, we were also interested in identifying novel molecular targets of IMP-1. Using in silico analysis of publicly available microarrays where IMP-1 was knocked down, we identified a panel of candidate target genes. Using qRTPCR and Western blot analysis, we then confirmed whether or not the mRNAs of candidate genes were decreased with IMP-1 knockdown and identified Protein Kinase C a (PKCa) as a new molecular target of IMP-1. Overall, this work has led to the identification and characterization of the first small molecule inhibitor of IMP-1 and has demonstrated that despite the fact that studies of the role of IMP-1 in cancer are rapidly expanding, there still remain novel molecular targets, such as PKCa, which may play critical roles in IMP-1 action in cancer cells.","abstract_has_math":false,"creators":["Mahapatra, Lily"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Molecular & Integrative Physi","degree_department":null,"school":null,"contributors":["Shapiro, David J.","Kemper, Jongsook Kim","Bolton, Eric","Anakk, Sayeepriyadarshini"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2016,"date_issued":"2016-07-07T19:52:38Z","date_published":"2016-07-07T19:52:38Z","updated_at":"2026-07-22T22:26:32Z","subjects":["IM-1","Small Molecule Inhibitor"],"languages":["en"],"rights":["Copyright 2016 Lily Mahapatra"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/90460","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Shapiro, David J.","Kemper, Jongsook Kim","Bolton, Eric","Anakk, Sayeepriyadarshini"]},{"key":"dc:creator","label":"Author","values":["Mahapatra, Lily"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2016-07-07T19:52:38Z","2016-01-25","2016-05"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Molecular & Integrative Physi"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["IM-1","Small Molecule Inhibitor"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2016 Lily Mahapatra"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/90460"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["RNA-binding proteins control a variety of biological processes ranging from messenger RNA splicing to transport and translation. These post-transcriptional events are critical for proper cell function. One emerging class of proteins functions in several of these capacities. The VICKZ family of RNA-binding proteins is involved in translation control, mRNA localization and mRNA stability. I have studied the Insulin-like Growth Factor-2 mRNA-Binding Protein 1 (IGF2BP1/IMP-1/CRD-BP). IMP-1 exhibits an oncofetal pattern of expression, where it is expressed in embryonic development and its expression is repressed shortly after birth. However, the IMP-1 gene is reactivated in many different human cancers. Overexpression of IMP-1 leads to increased levels of proteins that promote tumor growth, metastasis, and resistance to anticancer drugs and is associated with a poor prognosis. IMP-1 enhances proliferation and migration of cancer cells by binding to and stabilizing mRNAs important in cancer, such as c-Myc. Although the role of c-Myc in cancer has been well established, it has remained an elusive therapeutic target because of its role as a transcription factor in non-neoplastic proliferating cells. Given its oncofetal pattern of expression, targeting IMP-1 presents a novel approach to targeting c-Myc. To identify new chemical entities with therapeutic potential in IMP-1 positive cancer, we carried out a pilot screen using an in vitro fluorescence anisotropy microplate assay (FAMA) and found that this approach was robust and appropriate for high throughput screening. We then carried out a high throughput screen of approximately 150,000 small molecules. Reported here is BTYNB, the first small molecule inhibitor of IMP-1, BTYNB decreases levels of IMP-1 target mRNAs, inhibits proliferation of IMP-1 positive cancer cells, and functions through the unique mechanism of decreasing oncogene mRNA stability. We believe that BTYNB not only can be developed as a potential therapeutic agent, but also serves as a useful molecular tool, with which we can probe the actions of IMP-1 in cancer cells. In addition to identifying and characterizing the first small molecule inhibitor of IMP-1, we were also interested in identifying novel molecular targets of IMP-1. Using in silico analysis of publicly available microarrays where IMP-1 was knocked down, we identified a panel of candidate target genes. Using qRTPCR and Western blot analysis, we then confirmed whether or not the mRNAs of candidate genes were decreased with IMP-1 knockdown and identified Protein Kinase C a (PKCa) as a new molecular target of IMP-1. Overall, this work has led to the identification and characterization of the first small molecule inhibitor of IMP-1 and has demonstrated that despite the fact that studies of the role of IMP-1 in cancer are rapidly expanding, there still remain novel molecular targets, such as PKCa, which may play critical roles in IMP-1 action in cancer cells.","Submission original under an indefinite embargo labeled 'Open Access'. The submission was exported from vireo on 2016-07-07 without embargo terms","The student, Lily Mahapatra, accepted the attached license on 2016-01-13 at 17:41.","The student, Lily Mahapatra, submitted this Dissertation for approval on 2016-01-13 at 17:52.","This Dissertation was approved for publication on 2016-01-25 at 11:57.","DSpace SAF Submission Ingestion Package generated from Vireo submission #9035 on 2016-07-07 at 13:26:42","Made available in DSpace on 2016-07-07T19:52:38Z (GMT). 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One emerging class of proteins functions in several of these capacities. The VICKZ family of RNA-binding proteins is involved in translation control, mRNA localization and mRNA stability. I have studied the Insulin-like Growth Factor-2 mRNA-Binding Protein 1 (IGF2BP1/IMP-1/CRD-BP). IMP-1 exhibits an oncofetal pattern of expression, where it is expressed in embryonic development and its expression is repressed shortly after birth. However, the IMP-1 gene is reactivated in many different human cancers. Overexpression of IMP-1 leads to increased levels of proteins that promote tumor growth, metastasis, and resistance to anticancer drugs and is associated with a poor prognosis. IMP-1 enhances proliferation and migration of cancer cells by binding to and stabilizing mRNAs important in cancer, such as c-Myc. Although the role of c-Myc in cancer has been well established, it has remained an elusive therapeutic target because of its role as a transcription factor in non-neoplastic proliferating cells. Given its oncofetal pattern of expression, targeting IMP-1 presents a novel approach to targeting c-Myc. To identify new chemical entities with therapeutic potential in IMP-1 positive cancer, we carried out a pilot screen using an in vitro fluorescence anisotropy microplate assay (FAMA) and found that this approach was robust and appropriate for high throughput screening. We then carried out a high throughput screen of approximately 150,000 small molecules. Reported here is BTYNB, the first small molecule inhibitor of IMP-1, BTYNB decreases levels of IMP-1 target mRNAs, inhibits proliferation of IMP-1 positive cancer cells, and functions through the unique mechanism of decreasing oncogene mRNA stability. We believe that BTYNB not only can be developed as a potential therapeutic agent, but also serves as a useful molecular tool, with which we can probe the actions of IMP-1 in cancer cells. In addition to identifying and characterizing the first small molecule inhibitor of IMP-1, we were also interested in identifying novel molecular targets of IMP-1. Using in silico analysis of publicly available microarrays where IMP-1 was knocked down, we identified a panel of candidate target genes. Using qRTPCR and Western blot analysis, we then confirmed whether or not the mRNAs of candidate genes were decreased with IMP-1 knockdown and identified Protein Kinase C a (PKCa) as a new molecular target of IMP-1. Overall, this work has led to the identification and characterization of the first small molecule inhibitor of IMP-1 and has demonstrated that despite the fact that studies of the role of IMP-1 in cancer are rapidly expanding, there still remain novel molecular targets, such as PKCa, which may play critical roles in IMP-1 action in cancer cells.","Submission original under an indefinite embargo labeled 'Open Access'. The submission was exported from vireo on 2016-07-07 without embargo terms","The student, Lily Mahapatra, accepted the attached license on 2016-01-13 at 17:41.","The student, Lily Mahapatra, submitted this Dissertation for approval on 2016-01-13 at 17:52.","This Dissertation was approved for publication on 2016-01-25 at 11:57.","DSpace SAF Submission Ingestion Package generated from Vireo submission #9035 on 2016-07-07 at 13:26:42","Made available in DSpace on 2016-07-07T19:52:38Z (GMT). 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