{"id":{"repo_id":"umkc","oai_identifier":"oai:mospace.umsystem.edu:10355/97961"},"canonical_url":"https://search.dev.ndltd.org/etd/umkc/oai:mospace.umsystem.edu:10355/97961","repository":{"repo_id":"umkc","name":"University of Missouri - Kansas City","base_url":"https://mospace.umsystem.edu/oai/request"},"display":{"title":"Targeting extracellular matrix using siRNA for the treatment of liver fibrosis and pancreatic cancer","abstract":"The objective of this dissertation is to develop peptide-based siRNA delivery systems for the treatment of liver fibrosis and pancreatic ductal adenocarcinoma (PDAC). We recently discovered that silencing the poly (rC) binding protein 2 (PCBP2) gene in hepatic stellate cells (HSCs) and NIH 3T3 fibroblasts can reverse the accumulation of extracellular matrix in liver fibrosis and improve the penetration of small molecules in the tumor microenvironment, respectively. We previously developed a small peptide-based delivery system primarily composed of lysine and histidine residues, which improved the bioactivity and stability of siRNA. We, therefore, hypothesize that cholesteryl peptide/PCBP2 siRNA nanocomplexes modified with targeting and functional polyaminoacids/polypeptides could increase the specificity, delivery efficiency, and therapeutic efficacy. Furthermore, we also developed and evaluated a feasible rat model for alcoholic liver fibrosis. In Chapter 1, we briefly introduced the background of the dissertation research and presented the Statement of the Problems and Objectives. In Chapter 2, we reviewed the liver fibrogenesis and potential treatments for liver fibrosis. We also discussed the strategies to target desmoplastic stroma and cancer immunotherapy for pancreatic cancer. In Chapter 3, we presented the development of peptide-based siRNA nanocomplexes for HSC-specific drug delivery. We recently discovered a 12-mer ligand peptide for insulin-like growth factor 2 receptor (IGF2R) using the phage display technique, which significantly increased the binding affinity of nanoparticles to activated HSCs. In this chapter, we employed the IGF2R ligand peptide to improve the silencing efficiency and cellular uptake of the cholesteryl peptide/PCBP2 siRNA nanocomplexes. We incorporated glycine spacer and glutamate residues into the C-terminus of the dimeric IGF2R peptide ligands to non-covalently modify the surface of the siRNA nanocomplexes. We then compared the in vitro features of various IGF2R peptide-modified nanocomplexes. The resulting nanocomplexes demonstrated improved silencing efficiency activity, cellular uptake, and accumulation in the liver of rats with CCl₄-induced liver fibrosis. This strategy provides a promising platform for delivering antifibrotic siRNAs for liver fibrosis. In Chapter 4, we presented the development of a feasible rat model for alcoholic liver fibrosis and discussed its key characteristics. To replicate human drinking patterns, the rats were fed with a daily alcohol liquid diet and multiple alcohol binges, mimicking chronic and acute alcohol intake in humans, respectively. Carbon tetrachloride (CCl₄), the most commonly used hepatotoxin to induce liver fibrosis and cirrhosis in rodents, was administrated at a low dose to accelerate liver fibrogenesis. We discussed the pathophysiological features of liver injury and fibrosis induced by the combination of alcohol and low-dose CCl₄ treatment with various patterns. The results showed that the combination of alcohol and low-dose CCl₄ had a synergistic effect on increasing the levels of liver enzyme activity, collagen expression, HSCs activation, and cell proliferation. Consequently, this led to the progression of liver fibrosis. We successfully established an 8-week rat model of alcoholic liver fibrosis, providing a time-efficient model for future studies. In Chapter 5, we described a novel combination therapy approach for pancreatic ductal adenocarcinoma (PDAC). We developed a nanocarrier for delivering PCBP2 siRNA and an anti- programmed cell death ligand-1 (PD-L1) peptide simultaneously, aiming to improve the antitumor efficacy and delivery efficiency. Phosphorothioate (PS) linkage and 2’-O-methyl (2’-OMe) modified PCBP2 siRNA showed a longer half-life in mouse serum compared with unmodified PCBP2 siRNA. To increase the accumulation of siRNA and anti-PD-L1 in the tumor, the cholesteryl peptide/siRNA nanocomplexes was further condensed with plectin-1-G-pE6-M-PD-L1 (PGEMP) polypeptide. We assessed the characteristics of the resulting nanocomplexes, including particle size, cell viability, silencing activity, and serum stability. This strategy provides a promising platform for improving the anti-tumor efficiency of immunotherapy for PDAC.","abstract_html":"The objective of this dissertation is to develop peptide-based siRNA delivery systems for the treatment of liver fibrosis and pancreatic ductal adenocarcinoma (PDAC). We recently discovered that silencing the poly (rC) binding protein 2 (PCBP2) gene in hepatic stellate cells (HSCs) and NIH 3T3 fibroblasts can reverse the accumulation of extracellular matrix in liver fibrosis and improve the penetration of small molecules in the tumor microenvironment, respectively. We previously developed a small peptide-based delivery system primarily composed of lysine and histidine residues, which improved the bioactivity and stability of siRNA. We, therefore, hypothesize that cholesteryl peptide/PCBP2 siRNA nanocomplexes modified with targeting and functional polyaminoacids/polypeptides could increase the specificity, delivery efficiency, and therapeutic efficacy. Furthermore, we also developed and evaluated a feasible rat model for alcoholic liver fibrosis. In Chapter 1, we briefly introduced the background of the dissertation research and presented the Statement of the Problems and Objectives. In Chapter 2, we reviewed the liver fibrogenesis and potential treatments for liver fibrosis. We also discussed the strategies to target desmoplastic stroma and cancer immunotherapy for pancreatic cancer. In Chapter 3, we presented the development of peptide-based siRNA nanocomplexes for HSC-specific drug delivery. We recently discovered a 12-mer ligand peptide for insulin-like growth factor 2 receptor (IGF2R) using the phage display technique, which significantly increased the binding affinity of nanoparticles to activated HSCs. In this chapter, we employed the IGF2R ligand peptide to improve the silencing efficiency and cellular uptake of the cholesteryl peptide/PCBP2 siRNA nanocomplexes. We incorporated glycine spacer and glutamate residues into the C-terminus of the dimeric IGF2R peptide ligands to non-covalently modify the surface of the siRNA nanocomplexes. We then compared the in vitro features of various IGF2R peptide-modified nanocomplexes. The resulting nanocomplexes demonstrated improved silencing efficiency activity, cellular uptake, and accumulation in the liver of rats with CCl₄-induced liver fibrosis. This strategy provides a promising platform for delivering antifibrotic siRNAs for liver fibrosis. In Chapter 4, we presented the development of a feasible rat model for alcoholic liver fibrosis and discussed its key characteristics. To replicate human drinking patterns, the rats were fed with a daily alcohol liquid diet and multiple alcohol binges, mimicking chronic and acute alcohol intake in humans, respectively. Carbon tetrachloride (CCl₄), the most commonly used hepatotoxin to induce liver fibrosis and cirrhosis in rodents, was administrated at a low dose to accelerate liver fibrogenesis. We discussed the pathophysiological features of liver injury and fibrosis induced by the combination of alcohol and low-dose CCl₄ treatment with various patterns. The results showed that the combination of alcohol and low-dose CCl₄ had a synergistic effect on increasing the levels of liver enzyme activity, collagen expression, HSCs activation, and cell proliferation. Consequently, this led to the progression of liver fibrosis. We successfully established an 8-week rat model of alcoholic liver fibrosis, providing a time-efficient model for future studies. In Chapter 5, we described a novel combination therapy approach for pancreatic ductal adenocarcinoma (PDAC). We developed a nanocarrier for delivering PCBP2 siRNA and an anti- programmed cell death ligand-1 (PD-L1) peptide simultaneously, aiming to improve the antitumor efficacy and delivery efficiency. Phosphorothioate (PS) linkage and 2’-O-methyl (2’-OMe) modified PCBP2 siRNA showed a longer half-life in mouse serum compared with unmodified PCBP2 siRNA. To increase the accumulation of siRNA and anti-PD-L1 in the tumor, the cholesteryl peptide/siRNA nanocomplexes was further condensed with plectin-1-G-pE6-M-PD-L1 (PGEMP) polypeptide. We assessed the characteristics of the resulting nanocomplexes, including particle size, cell viability, silencing activity, and serum stability. This strategy provides a promising platform for improving the anti-tumor efficiency of immunotherapy for PDAC.","abstract_has_math":false,"creators":["Lin, Chien-Yu"],"institution":"University of Missouri--Kansas City","degree_name":"Ph.D. (Doctor of Philosophy)","degree_level":"Doctoral","degree_discipline":"Pharmaceutical Sciences (UMKC)","degree_department":null,"school":null,"contributors":[],"advisors":["Cheng, Kun (Professor)"],"committee_chairs":[],"committee_members":[],"year":2023,"date_issued":"2023","date_published":"2023","updated_at":"2026-07-24T05:18:08Z","subjects":[],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/10355/97961","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Cheng, Kun (Professor)"]},{"key":"dc:creator","label":"Author","values":["Lin, Chien-Yu"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2024-01-17T20:07:01Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2024-01-17T20:07:01Z"]},{"key":"dc:date.issued","label":"Date","values":["2023"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Pharmaceutical Sciences (UMKC)","Chemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Doctoral"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D. (Doctor of Philosophy)"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Missouri--Kansas City"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/10355/97961"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Title from PDF of title page, viewed January 3, 2025","Dissertation advisor: Kun Cheng","Vita","Includes bibliographical references (pages 104-135)","Dissertation (Ph.D.)--Division of Pharmaceutical Sciences, Department of Chemistry. University of Missouri--Kansas City, 2023"]},{"key":"dc:description.abstract","label":"Abstract","values":["The objective of this dissertation is to develop peptide-based siRNA delivery systems for the treatment of liver fibrosis and pancreatic ductal adenocarcinoma (PDAC). We recently discovered that silencing the poly (rC) binding protein 2 (PCBP2) gene in hepatic stellate cells (HSCs) and NIH 3T3 fibroblasts can reverse the accumulation of extracellular matrix in liver fibrosis and improve the penetration of small molecules in the tumor microenvironment, respectively. We previously developed a small peptide-based delivery system primarily composed of lysine and histidine residues, which improved the bioactivity and stability of siRNA. We, therefore, hypothesize that cholesteryl peptide/PCBP2 siRNA nanocomplexes modified with targeting and functional polyaminoacids/polypeptides could increase the specificity, delivery efficiency, and therapeutic efficacy. Furthermore, we also developed and evaluated a feasible rat model for alcoholic liver fibrosis. In Chapter 1, we briefly introduced the background of the dissertation research and presented the Statement of the Problems and Objectives. In Chapter 2, we reviewed the liver fibrogenesis and potential treatments for liver fibrosis. We also discussed the strategies to target desmoplastic stroma and cancer immunotherapy for pancreatic cancer. In Chapter 3, we presented the development of peptide-based siRNA nanocomplexes for HSC-specific drug delivery. We recently discovered a 12-mer ligand peptide for insulin-like growth factor 2 receptor (IGF2R) using the phage display technique, which significantly increased the binding affinity of nanoparticles to activated HSCs. In this chapter, we employed the IGF2R ligand peptide to improve the silencing efficiency and cellular uptake of the cholesteryl peptide/PCBP2 siRNA nanocomplexes. We incorporated glycine spacer and glutamate residues into the C-terminus of the dimeric IGF2R peptide ligands to non-covalently modify the surface of the siRNA nanocomplexes. We then compared the in vitro features of various IGF2R peptide-modified nanocomplexes. The resulting nanocomplexes demonstrated improved silencing efficiency activity, cellular uptake, and accumulation in the liver of rats with CCl₄-induced liver fibrosis. This strategy provides a promising platform for delivering antifibrotic siRNAs for liver fibrosis. In Chapter 4, we presented the development of a feasible rat model for alcoholic liver fibrosis and discussed its key characteristics. To replicate human drinking patterns, the rats were fed with a daily alcohol liquid diet and multiple alcohol binges, mimicking chronic and acute alcohol intake in humans, respectively. Carbon tetrachloride (CCl₄), the most commonly used hepatotoxin to induce liver fibrosis and cirrhosis in rodents, was administrated at a low dose to accelerate liver fibrogenesis. We discussed the pathophysiological features of liver injury and fibrosis induced by the combination of alcohol and low-dose CCl₄ treatment with various patterns. The results showed that the combination of alcohol and low-dose CCl₄ had a synergistic effect on increasing the levels of liver enzyme activity, collagen expression, HSCs activation, and cell proliferation. Consequently, this led to the progression of liver fibrosis. We successfully established an 8-week rat model of alcoholic liver fibrosis, providing a time-efficient model for future studies. In Chapter 5, we described a novel combination therapy approach for pancreatic ductal adenocarcinoma (PDAC). We developed a nanocarrier for delivering PCBP2 siRNA and an anti- programmed cell death ligand-1 (PD-L1) peptide simultaneously, aiming to improve the antitumor efficacy and delivery efficiency. Phosphorothioate (PS) linkage and 2’-O-methyl (2’-OMe) modified PCBP2 siRNA showed a longer half-life in mouse serum compared with unmodified PCBP2 siRNA. To increase the accumulation of siRNA and anti-PD-L1 in the tumor, the cholesteryl peptide/siRNA nanocomplexes was further condensed with plectin-1-G-pE6-M-PD-L1 (PGEMP) polypeptide. We assessed the characteristics of the resulting nanocomplexes, including particle size, cell viability, silencing activity, and serum stability. This strategy provides a promising platform for improving the anti-tumor efficiency of immunotherapy for PDAC."]},{"key":"dc:title","label":"Title","values":["Targeting extracellular matrix using siRNA for the treatment of liver fibrosis and pancreatic cancer"]}]}],"canonical_facts":{"dc:contributor.advisor":["Cheng, Kun (Professor)"],"dc:creator":["Lin, Chien-Yu"],"dc:date.accessioned":["2024-01-17T20:07:01Z"],"dc:date.available":["2024-01-17T20:07:01Z"],"dc:date.issued":["2023"],"dc:description":["Title from PDF of title page, viewed January 3, 2025","Dissertation advisor: Kun Cheng","Vita","Includes bibliographical references (pages 104-135)","Dissertation (Ph.D.)--Division of Pharmaceutical Sciences, Department of Chemistry. University of Missouri--Kansas City, 2023"],"dc:description.abstract":["The objective of this dissertation is to develop peptide-based siRNA delivery systems for the treatment of liver fibrosis and pancreatic ductal adenocarcinoma (PDAC). We recently discovered that silencing the poly (rC) binding protein 2 (PCBP2) gene in hepatic stellate cells (HSCs) and NIH 3T3 fibroblasts can reverse the accumulation of extracellular matrix in liver fibrosis and improve the penetration of small molecules in the tumor microenvironment, respectively. We previously developed a small peptide-based delivery system primarily composed of lysine and histidine residues, which improved the bioactivity and stability of siRNA. We, therefore, hypothesize that cholesteryl peptide/PCBP2 siRNA nanocomplexes modified with targeting and functional polyaminoacids/polypeptides could increase the specificity, delivery efficiency, and therapeutic efficacy. Furthermore, we also developed and evaluated a feasible rat model for alcoholic liver fibrosis. In Chapter 1, we briefly introduced the background of the dissertation research and presented the Statement of the Problems and Objectives. In Chapter 2, we reviewed the liver fibrogenesis and potential treatments for liver fibrosis. We also discussed the strategies to target desmoplastic stroma and cancer immunotherapy for pancreatic cancer. In Chapter 3, we presented the development of peptide-based siRNA nanocomplexes for HSC-specific drug delivery. We recently discovered a 12-mer ligand peptide for insulin-like growth factor 2 receptor (IGF2R) using the phage display technique, which significantly increased the binding affinity of nanoparticles to activated HSCs. In this chapter, we employed the IGF2R ligand peptide to improve the silencing efficiency and cellular uptake of the cholesteryl peptide/PCBP2 siRNA nanocomplexes. We incorporated glycine spacer and glutamate residues into the C-terminus of the dimeric IGF2R peptide ligands to non-covalently modify the surface of the siRNA nanocomplexes. We then compared the in vitro features of various IGF2R peptide-modified nanocomplexes. The resulting nanocomplexes demonstrated improved silencing efficiency activity, cellular uptake, and accumulation in the liver of rats with CCl₄-induced liver fibrosis. This strategy provides a promising platform for delivering antifibrotic siRNAs for liver fibrosis. In Chapter 4, we presented the development of a feasible rat model for alcoholic liver fibrosis and discussed its key characteristics. To replicate human drinking patterns, the rats were fed with a daily alcohol liquid diet and multiple alcohol binges, mimicking chronic and acute alcohol intake in humans, respectively. Carbon tetrachloride (CCl₄), the most commonly used hepatotoxin to induce liver fibrosis and cirrhosis in rodents, was administrated at a low dose to accelerate liver fibrogenesis. We discussed the pathophysiological features of liver injury and fibrosis induced by the combination of alcohol and low-dose CCl₄ treatment with various patterns. The results showed that the combination of alcohol and low-dose CCl₄ had a synergistic effect on increasing the levels of liver enzyme activity, collagen expression, HSCs activation, and cell proliferation. Consequently, this led to the progression of liver fibrosis. We successfully established an 8-week rat model of alcoholic liver fibrosis, providing a time-efficient model for future studies. In Chapter 5, we described a novel combination therapy approach for pancreatic ductal adenocarcinoma (PDAC). We developed a nanocarrier for delivering PCBP2 siRNA and an anti- programmed cell death ligand-1 (PD-L1) peptide simultaneously, aiming to improve the antitumor efficacy and delivery efficiency. Phosphorothioate (PS) linkage and 2’-O-methyl (2’-OMe) modified PCBP2 siRNA showed a longer half-life in mouse serum compared with unmodified PCBP2 siRNA. To increase the accumulation of siRNA and anti-PD-L1 in the tumor, the cholesteryl peptide/siRNA nanocomplexes was further condensed with plectin-1-G-pE6-M-PD-L1 (PGEMP) polypeptide. We assessed the characteristics of the resulting nanocomplexes, including particle size, cell viability, silencing activity, and serum stability. This strategy provides a promising platform for improving the anti-tumor efficiency of immunotherapy for PDAC."],"dc:identifier.uri":["https://hdl.handle.net/10355/97961"],"dc:title":["Targeting extracellular matrix using siRNA for the treatment of liver fibrosis and pancreatic cancer"],"thesis:degree_discipline":["Pharmaceutical Sciences (UMKC)","Chemistry"],"thesis:degree_level":["Doctoral"],"thesis:degree_name":["Ph.D. (Doctor of Philosophy)"],"thesis:institution_name":["University of Missouri--Kansas City"]},"updated_at":"2026-07-24T05:18:08Z"}