Targeting PI3K/BRD4 and Hedgehog Pathway in Alcohol Associated Liver Disease

靶向 PI3K/BRD4 和 Hedgehog 通路治疗酒精相关性肝病

• 批准号: 10351877
• 负责人: Virender Kumar
• 金额: $ 10.56万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10351877
• 关键词: AKT inhibition; Acetaldehyde; Alcohol abuse; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Area; Award; Binding; Biodistribution; Blood; CYP2E1 gene; Cell Survival; Cells; Cessation of life; Cirrhosis; Cleaved cell; Collaborations; DNA Damage; Data; Deposition; Development; Diagnosis; Diet; Disease; Disease Progression; Disease regression; Drug Delivery Systems; Drug Kinetics; Encapsulated; Environment; Erinaceidae; Ethanol; Extracellular Matrix; Faculty; Fatty Acids; Fatty acid glycerol esters; Fibroblasts; Fibrosis; Funding; GLI Family Protein; GLI gene; Gene Family; Genes; Goals; Grant; HMGB1 Protein; Hepatic Stellate Cell; Hepatitis C Therapy; Hepatocyte; Human; Hydrophobicity; Immune response; In Vitro; Industry; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Knowledge; Kupffer Cells; Leadership; Leaky Gut; Ligands; Lipid Peroxidation; Lipids; Lipopolysaccharides; Liver; Liver Fibrosis; Liver diseases; Malignant neoplasm of pancreas; Mediating; Mentors; Morbidity - disease rate; Mus; NF-kappa B; Nature; Necrosis; Oxides; PI3K/AKT; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor beta Receptor; Production; Protein-Serine-Threonine Kinases; Proteins; Proto-Oncogene Proteins c-akt; Public Health; Pulmonary Inflammation; Recombinants; Research; Research Personnel; Role; Sampling; Scientist; Signal Pathway; Signal Transduction; Students; Surface; TLR4 gene; Training; Transcription Coactivator; Treatment Efficacy; United States; Up-Regulation; Writing; alcohol abstinence; alcohol abuse therapy; amphiphilicity; base; c-myc Genes; career; career development; cell transformation; chronic liver injury; conventional therapy; cytokine; dietary control; experience; fibroblast-activating factor; gene function; in vivo; inhibitor/antagonist; innovation; kidney fibrosis; liver inflammation; liver injury; liver transplantation; member; nanocarrier; nanomedicine; nanoparticle; new therapeutic target; next generation; novel; novel therapeutics; overexpression; prevent; protein expression; response; skills; smoothened signaling pathway; tenure track; wound healing

PROJECT SUMMARY This K01 application aims to promote the applicant's development to become a multi-disciplinarily trained and independent academic researcher. This application's collective strengths are defined in these 3 major areas: 1) Credentials: PI's application builds upon his productive track-record to achieve scientific independence in the field of drug delivery and liver fibrosis, including alcohol-associated liver disease (AALD). The PI's goals include enhancing grantsmanship, leadership, and team management skills to become an independent, tenure-track academic researcher. In addition to technical training, PI intends to build skills in communicating with faculty members, industry members, and students. 2) Training Environment: Drs. Mahato (mentor) and Kharbanda, and Cheng (co-mentors) are world-class researchers and fully committed to advancing the PI's career. Particular emphasis will be given to grant writing, collaboration building, and applying for external funding. The mentoring committee has strong credentials in developing the next generation of successful academic scientists. The knowledge and experience gained during this award period will help the candidate generate data to apply for an R21/R01 grant. 3) Innovative Research: PI's central hypothesis is that alcohol-induced activation of inflammatory pathways is mediated by AKT and BRD4 and synergize with the hedgehog (Hh) pathway to promote AALD. Simultaneous inhibition of these pathways using targeted nanomedicine could alleviate the progression of AALD and related morbidities. The PI has generated the following preliminary results; (a) Conditioned media from the EtOH treated primary human hepatocytes significantly stimulated hepatic stellate cells (HSCs) as determined by α-SMA expression levels and upregulated GLI1/2 activity; (b) The protein cMYC is upregulated in patient samples diagnosed with alcoholic hepatitis; (c) Treatment with novel inhibitors MDB5 (Hh) and SF2523 (PI3K/BRD4) showed decreased their target genes in HSC-T6 cells; (d) EtOH diet-fed mice show increased p- AKT, cMYC, and GLI1/2 protein levels in the liver compared to mice on the control diet; (e) Treatment with MDB5 and SF2523 decreased liver injury markers ALT and AST, and lowered GLI1, cMYC, and p-AKT protein levels in ethanol-fed mice; (f) Novel cleavable amphiphilic peptide (CAP) was self-assembled into nanoparticles (NPs) of size range 100± 10 nm loaded with both the drugs (payload 5% w/w); (g) CAP-NPs were sensitive to fibroblast activation protein (FAP-α), and NPs dissembled in presence of recombinant FAP-α. Based on these robust results, the proposal has the following specific aims: Aim 1. Establish the role of AKT/BRD4 dual inhibitor SF2523 and Hh inhibitor MDB5 in AALD. Aim 2. Formulate and characterize HSC targeted CAP-NP loaded with SF2523 and MDB5. Aim 3. Determine therapeutic efficacy of SF2523 and MDB5 loaded pPB-CAP-NPs in ALD mice. The new knowledge and nanomedicine generated in this proposal may open new therapeutic avenues for the treatment of AALD.

项目总结 这份K01申请书旨在促进申请者的发展，使其成为一名受过多学科训练和 独立的学术研究员。此应用程序的整体优势在以下三个主要方面进行了定义：1) 证书：Pi的应用程序建立在他富有成效的记录基础上，以实现科学独立 药物输送和肝纤维化领域，包括酒精相关性肝病(AALD)。私家侦探的目标包括 加强团队精神、领导力和团队管理技能，以成为独立的终身教职轨道 学术研究人员。除了技术培训外，PI还打算培养与教职员工沟通的技能 成员、行业成员和学生。2)培训环境：马哈托博士(导师)和坎潘达博士，以及 程(共同导师)是世界级的研究人员，完全致力于推动PI的事业发展。特例 重点将放在赠款的撰写、合作建设和申请外部资金上。指导工作 委员会在培养下一代成功的学术科学家方面拥有很强的资历。这个 在此授奖期间获得的知识和经验将帮助候选人生成数据以申请 R21/R01赠款。3)创新研究：Pi的中心假说是酒精诱导炎症的激活 AKT和BRD4是AKT和BRD4介导的信号转导通路，并与HH通路协同促进AALD的发生。 使用靶向纳米药物同时抑制这些通路可以减缓AALD的进展 以及相关的疾病。PI产生了以下初步结果；(A)来自 乙醇处理的原代人肝细胞显著刺激肝星状细胞(HSCs) α-SMA表达水平和GLI1/2活性上调；(B)患者cMYC蛋白上调 诊断为酒精性肝炎的样本；(C)新型抑制剂MDB5(HH)和SF2523的治疗 在HSC-T6细胞中，(PI3K/BRD4)显示其靶基因减少；(D)Etoh饮食喂养的小鼠显示其靶基因表达增加。 肝脏AKT、cMYC和GLI1/2蛋白水平与对照组小鼠的比较；(E) MDB5和SF2523可降低肝损伤标志物ALT和AST，降低GLI1、cMYC和p-AKT蛋白 (F)新型可裂解两亲性多肽(CAP)自组装成纳米颗粒 (G)CAP-NPs对药物敏感 成纤维细胞激活蛋白(FAP-α)，以及在重组FAP-α存在下的NPs。基于这些 结果稳健，该提案有以下具体目标：目的1.确定AKT/BRD4双重抑制物的作用 SF2523和HH抑制剂MDB5在AALD中的表达。目的2.制备和鉴定负载CAP-NP的HSC靶向 使用SF2523和MDB5。目的3.检测SF2523和MDB5负载ppb-CAP-NPs对人肺癌的治疗作用 ALD小鼠。在这项提议中产生的新知识和纳米医学可能会打开新的治疗途径 治疗急性酒精性肝病。