项目来源

美国卫生和人类服务部基金(HHS)

项目主持人

ORSBURN,BENJAMIN CARL

项目受资助机构

JOHNS HOPKINS UNIVERSITY

立项年度

2022

立项时间

未公开

项目编号

5R01GM10385310

研究期限

未知 / 未知

项目级别

国家级

受资助金额

360250.00美元

学科

INTERNAL MEDICINE/MEDICINE

学科代码

未公开

基金类别

Research Projects

Acute Liver Failure ; Address ; Adverse event ; Anti-Retroviral Agents ; Antiepileptic Agents ; Apoptosis ; Area ; BIM Bcl-2-binding protein ; Biological Assay ; CRISPR/Cas technology ; Carbamazepine ; Cell Death ; Cells ; Cellular Stress ; Cessation of life ; Chemicals ; Clinical ; Clinical Data ; Clinical Markers ; Complex ; Cyclic AMP-Dependent Protein Kinases ; Data ; Diclofenac ; Endoribonucleases ; Enzymes ; Event ; Foundations ; Genes ; Genetic ; Genetic Transcription ; Genetic Variation ; Genomic DNA ; Genotype ; Goals ; Hepatic ; Hepatocyte ; Histology

参与者

未公开

参与机构

未公开

项目标书摘要：Drug-induced hepatotoxicity is a leading cause of both the withdrawal of approved drugs fromthe market and the attrition of new chemical entities during the drug development process;however,the mechanisms underlying drug-induced hepatotoxicity are not fully understood.Wehave used efavirenz,an antiretroviral drug that is hepatotoxic in certain patients,as a modelcompound to investigate cellular signaling mechanisms that may play a causal role in drug-induced hepatocyte death.Previously,using primary human hepatocytes,we demonstrated thatefavirenz and the major oxidative metabolite of efavirenz,denoted as 8-hydroxyefavirenz(8-OHefavirenz),stimulate cell death in a manner that is dependent upon activation of the stresskinase c-Jun N-terminal kinase and upregulation of the proapoptotic protein BimEL(Bcl-2interacting mediator of cell death extra long).Subsequently,we have reported that efavirenz canalso activate inositol requiring enzyme 1α(IRE1α),a key regulator of cell stress that lies upstreamof JNK and BimEL.The goal of this proposal is to determine the mechanism by which efavirenzand 8-OHefavirenz activate BimEL and IRE1α,while also gaining a mechanistic understanding ofhow genetic variation in IRE1α might impact efavirenz and 8-OHefavirenz-induced cell death.Importantly,we will leverage the insights we have gained through using efavirenz as a modelcompound and employ prototypic hepatotoxic drugs beyond efavirenz,namely carbamazepine,diclofenac and isoniazid,in order to establish BimEL and IRE1α as central regulators of drug-induced hepatotoxicity across a range of drug classes.The aims are as follows:(1)to test thehypothesis that BimEL acts as an executioner of cell death in response to efavirenz and otherprototypic hepatotoxic drugs:BimEL null mice will be used to determine whether the absence ofBimEL prevents hepatotoxicity stimulated by the hepatotoxic drugs being investigated here;CRISPR/Cas9 systems will be used to determine the role of effector proteins,Bax and Bak,thatare downstream of BimEL in modulating hepatocyte death;CRISPR/Cas9 and reporter geneassays will be used to define the mechanism by which efavirenz,8-OHefavirenz and otherhepatotoxic drugs regulate the transcription of BimEL;efavirenz analogs will be employed in orderto elucidate the structure-activity relationship of BimEL activation by efavirenz;(2)to test thehypothesis that IRE1α is a central upstream regulator of drug-induced hepatotoxicity that isstimulated by several classes of drugs:we will determine whether efavirenz,8-OHefavirenz,andother hepatotoxic drugs stimulate formation of the IRE1α/TRAF2/ASK1/JNK complex that resultsin IRE1α-dependent activation of JNK;we will test the impact of naturally occurring geneticvariants of IRE1α on activity and cell death.It is expected that these studies will define BimELand IRE1α activation as important molecular mechanisms by which a range of drugs induce-hepatotoxicity.

项目官员

KODURI,SAILAJA

项目持续时间

10 years

项目负责机构类型

SCHOOLS OF MEDICINE