Biomarkers to Measure Treatment Response for Alcohol Dependence

描述（由申请人提供）：K23申请的科学目标是开发一种新的基因组生物标志物组合（5 '-HTTLPR-LL基因型携带者中的血清素转运蛋白基因（SERT）mRNA表达水平），用于定量预测近期饮酒水平。该研究的假设来自候选人的一系列初步工作，该工作首先使用生化和药物遗传学研究证明了基于52-HTTLPR等位基因的饮酒严重程度差异;扩展这些发现，下一组初步实验提供了SERT mRNA表达水平与LL基因型携带者回顾性收集的自我报告的饮酒量之间正相关的证据。为了前瞻性验证5 '-HTTLPR-LL基因型和SERT mRNA表达水平的生物标志物组合，我提出了在5年K23期间分两个阶段进行的一系列实验。在第一阶段，我会招募一批成年人， 具有相同比例的LL和LS/SS基因型的男性和女性重度社交饮酒者：在人类实验室中，采用3 × 3平衡拉丁方设计的顺序和序列，对两个基因型组施用酒精。在人类实验室实验期间每天从参与者收集样本，我将进行SERT基因表达测定以评估所提出的生物标志物的有效性和特异性（具体目标I），并研究响应于交替剂量的酒精的全基因组microRNA表达变化，以鉴定调节基于5- HTTLPR等位基因的SERT基因转录水平的microRNA（具体目标II）。具体目标二也将有助于确定新的标记大量饮酒 这在具体目标I未实现的情况下尤其重要。只有在成功验证生物标志物后，才进行第二阶段，以测试补充目标。在阶段II中，我将比较所提出的标志物与%CDT和EtG的特异性（补充目的I），并测试SERT生物标志物组合的灵敏度，以检测在治疗临床试验中登记的酒精依赖个体的存档样品中的饮酒量（补充目的II）。如果得到验证，我相信拟议的SERT生物标志物组合可能有助于开发有效的药物来治疗饮酒问题，这是美国环境中第三大生活方式相关的死亡原因：我在弗吉尼亚大学精神病学和神经行为科学系的环境非常支持进行这项K23奖。我们的部门致力于在临床，转化和基础科学研究方面进行高质量的多学科研究。目前，我在神经生物学研究部担任初级教员，有超过15名NIH资助的教员研究人员，专注于各种成瘾的遗传、生物、药理和行为方面。我将在整个K3奖期间使用的三个设施，弗吉尼亚大学成瘾研究和教育中心（UVa CARE），弗吉尼亚大学临床精神药理学研究单位（CPRU）和神经生物学实验室，都在我们的部门内。我在这个项目上的主要导师是Bankole A教授。约翰逊，谁是药物开发领域的领导者，用于治疗成瘾，并已成功地指导许多初级研究人员在过去几年中有前途的学术生涯。此外，我在获奖期间的培训将得到一个完善的共同导师团队的加强，其中包括Ming D教授。为了进一步提高我在成瘾遗传学方面的技能和知识，Nassima Ait-Daoud博士定期就参与者相关问题进行咨询，Dr.生物统计学顾问马先生和高级生物统计学家王新群先生负责数据分析相关问题。通过我们在NIH资助的赠款，共同撰写论文以及他们对K23应用程序开发的支持，我与导师进行富有成效的合作的潜力是显而易见的。候选人：我是一名医生科学家，对研究酒精依赖的分子和遗传机制以及个人对治疗的个人反应感兴趣。我过去在遗传学和生物化学领域的研究训练为我在酒精成瘾的基础科学研究方面提供了良好的基础。有了K23提案的奖励，我将能够在受控条件下进行酒精使用模拟的人类实验室范例方面发展更多的专业知识，为我提供了独特的机会来发现如何将我在培训期间学到的分子和生化技术与人类精神药理学干预相结合。通过在拟议的K23奖期间利用这些干预措施，我将实现我的短期职业目标，即开发一种准确的生物标志物来测量酒精消费水平的瞬时变化，解决酒精成瘾治疗领域的关键需求。我的总体职业目标是通过靶向酒精成瘾的分子遗传机制来改善酒精成瘾的治疗，以确定药物治疗的新方向。在拟议的五年结束时，通过我从指导研究项目中获得的经验和知识，重点是人类心理药理学干预，再加上正式的课程工作，研讨会，参加国家和国际科学会议，我将成为一名成功的独立翻译研究科学家，专注于分子心理药理学。

DESCRIPTION (provided by applicant): The scientific goal of the K23 application is to develop a novel genomic biomarker combination (Serotonin transporter gene (SERT) mRNA expression levels in 5'-HTTLPR-LL genotype carriers) for quantitatively predict recent alcohol consumption levels. The hypothesis for the study was derived from a series of preliminary work by the candidate that first demonstrated a 52-HTTLPR allele based difference in drinking severity using biochemical and pharmacogenetic studies; extending these findings, the next set of preliminary experiments provided evidence for a positive association between SERT mRNA expression levels and retrospectively collected self-reported alcohol consumption amounts, in LL genotype carriers. To prospectively validate the biomarker combination of 5'-HTTLPR-LL genotype and SERT mRNA expression levels, I propose a series of experimentations conducted in two stages during the 5-year K23 period. In stage I, I will recruit a cohort of adult, male and female heavy social drinkers with LL and LS/SS genotypes in equal proportions: Alcohol will be administered to both genotype groups employing a 3x3 Diagram balanced Latin square design for order and sequence in a human laboratory. With the samples collected daily from the participants during the human laboratory experiments, I will perform SERT gene expression assays to assess validity and specificity of the proposed biomarker (Specific aim I),and study genome-wide microRNA expression alterations in response to alternating doses of alcohol to identify microRNAs that regulate 5¿- HTTLPR allele-based SERT gene transcription levels (Specific aim II). Specific aim II would also help identify novel markers for heavy drinking which is particularly important in case specific aim I is not achieved. Stage II will be performed to test the supplementary aims only upon successful validation of the biomarker. In stage II, I will compare the specificity of the proposed marker with %CDT and EtG (Supplementary aim I) and test the Sensitivity of the SERT biomarker combination to detect amounts of drinking, in archived samples from alcohol dependent individuals, enrolled in treatment clinical trials (Supplementary aim II). If validated, I believe the proposed SERT biomarker combination may aid in developing efficacious medications to treat problem drinking, which is the third leading life-style related cause of death in the U.S. Environment: My environment at the University of Virginia, Department of Psychiatry and Neurobehavioral Sciences is highly supportive for conducting this K23 award. Our department is well committed and recognized for conducting high quality multi-disciplinary research in clinical, translational and basic sciences research. Currently I serve as a junior faculty member at the Division of Neurobiological Studies with over 15 NIH-funded faculty researchers focusing on genetic, biological, pharmacological, and behavioral aspects of various addictions. The three facilities that I will utilize throughout the K3 award period, University of Virginia Center for Addiction Research and Education (UVa CARE), University of Virginia clinical Psychopharmacology research unit (CPRU), and the Neurobiology laboratory, are all housed within our division. My primary mentor on this project is Professor Bankole A. Johnson, who is a leader in the field of medications development for the treatment of addictions and has successfully mentored many junior investigators throughout the past several years to have promising academic careers. Additionally, my training during the award period will be enhanced by a well-established team of co-mentors, including Professor Ming D. Li to further my skills and knowledge in addiction genetics, Dr. Nassima Ait-Daoud with regular consultations on participant related issues, Dr. Jennie Z. Ma on Biostatistical consultations and Senior Biostatistician Xin Qun Wang with data analyses related issues. My potential for productive collaboration with my mentors is evident through our work on NIH funded grants, coauthoring papers, and their support in the development of this K23 application. Candidate: I am a physician scientist interested in studying molecular and genetic mechanisms underlying alcohol dependence and an individuals' personal response to treatment. My past research training in genetic and biochemistry fields have provided me with an excellent foundation in basic sciences research in alcohol addiction. With the award of this K23 proposal, I would be able to develop additional expertise in conducting human laboratory paradigms of alcohol use modeled under controlled conditions, providing me with the unique opportunity to discover how to integrate molecular and biochemical techniques that I have learned during my training, with human psychopharmacological interventions. By utilizing these interventions during the proposed K23 award period, I will realize my short-term career goal of developing an accurate biomarker to measure transient changes in alcohol consumption levels, addressing a critical need in the field of alcohol addiction treatment. My overarching career goal is to improve treatment for alcohol addiction, through targeting molecular genetic mechanisms underlying alcohol addiction to identify new directions in pharmacotherapy. At the end of the proposed five years, through the experience and knowledge that I gain from mentored research projects focusing on human psychopharmacological interventions, coupled with formal course work, seminars, participation in national and international scientific conferences, I will emerge a successful independent translational research scientist with a focus on molecular psychopharmacology.