Biomarkers to Measure Treatment Response for Alcohol Dependence

描述(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记物组合(5‘-HTTLPR-LL型携带者的5’-羟色胺转运体基因(SERT)mRNA表达水平)，以定量预测最近的酒精消费水平。这项研究的假设来自候选人的一系列初步工作，这些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；扩展了这些发现，下一组初步实验提供了证据，证明在LL基因携带者中，SERT mRNA表达水平与回顾收集的自我报告的饮酒量之间存在正相关。为了前瞻性地验证5‘-HTTLPR-LL基因和SERT mRNA表达水平的生物标志物组合，我提议在5年的K23期间分两个阶段进行一系列实验。在第一阶段，我将招募一群成年人， 男性和女性重度社交饮酒者的LL和LS/SS基因型比例相等：在人体实验室中，将采用3x3图平衡拉丁方设计的顺序和顺序，对这两个基因型组给予酒精。利用在人体实验室实验期间每天从参与者那里收集的样本，我将进行SERT基因表达分析，以评估所提出的生物标记物的有效性和特异性(特定目标I)，并研究改变剂量的酒精对全基因组microRNA表达的影响，以识别调节基于�-HTTLPR等位基因的SERT基因转录水平的microRNA(特定目标II)。Special Aim II还将有助于确定酗酒的新标记物 这在具体目标I没有实现的情况下尤其重要。只有在生物标记物成功验证后，才会执行第二阶段来测试补充目标。在第二阶段，我将比较建议的标记物与%CDT和EtG(补充目标I)的特异性，并测试SERT生物标记物组合的敏感性，以检测参加治疗临床试验(补充目标II)的酒精依赖者的存档样本中的饮酒量。如果得到验证，我相信建议的SERT生物标记物组合可能有助于开发有效的药物来治疗问题饮酒，这是美国环境中与生活方式相关的第三大死因：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举办这个K23奖项。我们系致力于在临床、翻译和基础科学研究方面进行高质量的多学科研究，并得到了认可。目前，我在神经生物学研究部担任初级教员，有超过15名由NIH资助的教员研究人员，专注于各种成瘾的遗传、生物学、药理学和行为方面的研究。在整个K3颁奖期间，我将使用的三个设施，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，都在我们部门内。我在这个项目上的主要导师是班科尔·A·约翰逊教授，他是治疗成瘾的药物开发领域的领导者，在过去几年里成功地指导了许多初级调查人员，使他们在学术生涯中拥有光明的前景。此外，在颁奖期内，我的培训将由一支完善的联合导师团队加强，包括提升我在成瘾遗传学方面的技能和知识的李明达教授、定期咨询参与者相关问题的Nassima Ait-Daoud博士、生物统计咨询的Jennie Z.Ma博士以及与数据分析相关的高级生物统计师王新群。通过我们在NIH资助的拨款、共同撰写的论文以及他们对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显而易见。候选人：我是一名内科科学家，研究酒精依赖的分子和遗传机制，以及个人对治疗的反应。我过去在遗传和生物化学领域的研究培训为我在酒精成瘾的基础科学研究方面提供了良好的基础。有了这项K23计划的奖励，我将能够在控制条件下进行酒精使用的人体实验室范例方面开发更多的专业知识，为我提供一个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获奖期内利用这些干预措施，我将实现我的短期职业目标，即开发一种准确的生物标记物来测量酒精消费水平的瞬时变化，满足酒精成瘾治疗领域的迫切需求。我的首要职业目标是改善酒精成瘾的治疗，通过靶向酒精成瘾的分子遗传机制来确定药物治疗的新方向。在拟议的五年结束时，通过我从专注于人类精神药理学干预的指导研究项目中获得的经验和知识，再加上正规的课程工作、研讨会、参加国内和国际科学会议，我将成为一名成功的独立翻译研究科学家，专注于分子精神药理学。

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.