GENETIC DERMINATION OF PPH EXPRESSION

原发性肺动脉高压(PPH)是一种小肺动脉疾病，可以是家族性的(FPPH)或 零星的。骨形态发生蛋白受体2(BMPR2)基因突变是导致这两种疾病的主要原因 FPPH和散发性PPH。据报道，BMPR2基因外显子的异质性突变在~50%和 家族性病例和散发性病例分别占25%。其他人使用负荷超声心动图获得连锁数据 这表明，附近的第二个基因座也可能导致胎膜早破。FPPH在我国的渗透率仅为~20% 家人。对FPPH肺部解剖病变的研究显示克隆性，表明杂合性丢失 (LOH)和/或体细胞突变，这可能表明两次击中模式是降低外显率的基础 FPPH。而多个FPPH家系表现出遗传预期(连续的较早发病 世代)，没有报道BMPR2等位基因的变化来解释预期。我们假设1) BMPR2的等位基因异质性导致了许多FPPH和BMPR2突变的散发病例 2)核或线粒体修饰基因变异，杂合性缺失和/或获得性体细胞 3)BMPR2或修饰性基因的变异导致FPH的外显率降低 (4)基因座的异质性可能是导致FPPH的原因之一。为了更好地 了解PPH的分子基础，我们将确定BMPR2中存在的等位基因异质性 不同的FPPH家系和散发性病例的等位基因，识别和表征修饰基因或 BMPR2中影响FPPH外显率的体细胞变化，确定可能导致 并确定我们的家庭中是否存在基因座异质性(基因座异质性将是 在项目1中审查)。我们的研究具有普遍的意义，因为它们将为发病机制提供洞察力。 一种重要的常染色体显性遗传病，表现出可变的表达，外显性降低和 期待。 范德比尔特大学医学中心 儿科医学遗传学分会 DD-2205北区医疗中心 田纳西州纳什维尔，邮编：37232-2578 关键人员。请参阅说明。使用延续 调查员。按字母顺序列出所有其他关键人员 名字 菲利普斯，约翰·A，嗨，医学博士。 加迪帕蒂，拉迪卡，M.B.B.S. 科克·B·莱恩，博士。 Vnencak-Jones，Cindy，博士 根据需要提供所需信息的页面 请按顺序，姓氏在前。 组织 范德比尔特大学医学中心 范德比尔特大学医学中心 范德比尔特大学医学中心 范德比尔特大学医学中心 小灵通398(05/01版)_第19 8页 在整个申请过程中，在底部连续编号，不使用3a、3b等就足够了。 格式如下所示。从本金开始 在项目中的角色 项目负责人 调查员 调查员 调查员 表单第2页 O项目III：PPH表达的遗传测定 首席调查员/项目主任(最后、第一、中间)：Loyd I James E.V.M.D. 从头到尾 初步预算期明细预算 直接成本仅为7/1/2003 6/30/2004 人员(仅适用于申请者组织)请求的金额百分比(省略美分) 键入Effort Inst. APPT在基本工资福利上的名称角色 项目(月)项目。申请薪资BENEFTTS合计 本金 约翰·菲利普斯医学博士1225%166,700 41,675 6,626 48,301 调查员 Kirk B.Lane博士调查员12 30%93,600 28,080 6,318 34,398 [Cindy Vnencak-]One，博士调查员12.5%109,101 5,455 1,227 6,682 黛博拉·默多克博士调查员12 10%75,400 i 7,540 1,697 9,237 梅丽莎·普林斯实验室经理12 30%45,125 13,538 3,466 17,004 拉迪卡·加迪帕蒂12 25%46,188 11,547 2,956 14,503 托马斯·布莱克威尔实验室经理12 25%40,372 10,093 2,584 12,677 Res Asst III 12 100%37,500 37,500 9,600 47,100 Kuisok Keel技术员12 5%30,623 1,531 392 1,923 Res Asst II 12 100%32,500 32,500 8,320 40,820 小计189,459 43,186 232,645 顾问费 0 设备(分项) 0 用品(按类别分项列出) 7,000项预期研究的基因分型 27,000项BMPR2研究的基因分型和测序 陆恭蕙研习用品400 南方吸墨用品600份 组织培养用品15,000份 分子生物学用品(一次性用品、塑料制品、玻璃器皿、吸管等)14,500件 普通实验室用品11,000件75,500件 旅行 0 病人护理费用为0 住院病人 门诊部0 改建和翻新(按类别分列) 无%0 其他费用(按类别分项列出) 软件升级1,000 Lf000 初步预算期间的直接费用小计30E，1451美元 0 联合体/合同直接成本 成本设施和管理成本%0 初步预算期(Item7a，FacePage)的直接费用总额为309,1451美元 仅限SBIR/STTR：要求固定费用 PHS398(05/01版)(第19页)表格第4页 在整个应用程序底部连续编号页码。不要使用3a、3b等字样。

Primary pulmonary hypertension (PPH) is a disease of small pulmonary arteries which can be familial (FPPH) or sporadic. Mutations in the bone morphogenic protein receptor 2 (BMPR2) gene cause a major proportion of both FPPH and sporadic PPH. Heterogeneous mutations have been reported in the exons of BMPR2 in ~50% and ~25% of familial and sporadic cases, respectively. Using stress echocardiography others obtained linkage data suggesting that a second, nearby locus may also cause FPPH. The penetrance of FPPH is only ~20% in our families. Studies of dissected lesions from FPPH lungs show clonality suggesting that loss of heterozygosity (LOH) and/or somatic mutations which may indicate that a two hit model underlies the reduced penetrance of FPPH. While multiple FPPH families exhibit genetic anticipation (earlier onset of disease in successive generations), no BMPR2 allelic changes have been reported that explain anticipation. We hypothesize that 1) allelic heterogeneity in BMPR2 causes many of the FPPH and sporadic cases in which BMPR2 mutations have not been identified, 2) variation in nuclear or mitochondrial modifier genes, LOH and/or acquired somatic imutations contribute to the reduced penetrance of FPPH, 3) variations in BMPR2 or a modifier gene cause the anticipation seen in FPPH, and 4) locus heterogeneity may contribute to the etiology of FPPH. To better understand the molecular basis of PPH we will determine the allelic heterogeneity that exists in the BMPR2 alleles of various FPPH kindreds and sporadic cases, identify and characterize variations in modifier genes or somatic changes in BMPR2 that affect the penetrance of FPPH, identify genetic alterations that may cause anticipation in FPPH and determine if locus heterogeneity exists in our families (locus heterogeneity will be examined in Project 1). Our studies are of general interest because they will provide insight to the pathogenesis of an important autosomal dominant disease that exhibits variable expression, reduced penetrance and anticipation. Vanderbilt University Medical Center Division of Pediatric Medical Genetics DD-2205 Medical Center North Nashville, TN 37232-2578 KEY PERSONNEL. See instructions. Use continuation Investigator. List all other key personnel in alphabetical Name Phillips, John A., HI, M.D. Gaddipatti, Radhika, M.B.B.S. Lane, Kirk B., Ph.D. Vnencak-Jones, Cindy, Ph.D. pages as needed to provide the required information order, last name first. Organization Vanderbilt University Medical Center Vanderbilt University Medical Center Vanderbilt University Medical Center Vanderbilt University Medical Center PHS 398 (Rev. 05/01) _ Page 19 8 Number pages consecutively at the bottom throughout the application, uo not use suffices such as 3a, 3b. in the format shown below. Start with Principal Role on Project Project Leader Investigator Investigator Investigator Form Page 2 O Project III: Genetic Determination of PPH Expression Principal Investigator/Program Director (Last, first, middle): Loyd I James E.v M.D. FROM THROUGH DETAILED BUDGET FOR INITIAL BUDGET PERIOD DIRECT COSTS ONLY 7/1/2003 6/30/2004 PERSONNEL (Applicant orgam'zabbn only) % DOLLAR AMOUNT REQUESTED(omit cents) TYPE EFFORT INST. NAME ROLE ON APPT ON BASE SALARY FRINGE PROJECT (months) PROJ. SALARY REQUESTED BENEFTTS TOTALS Principal John Phillips, M.D. 12 25% 166,700 41,675 6,626 48,301 Investigator Kirk B. Lane, Ph.D. Investigator 12 30% 93,600 28,080 6,318 34,398 Cindy Vnencak-]ones, Ph.D. Investigator 12 5% 109,101 5,455 1,227 6,682 Deborah Murdock Ph.D. Investigator 12 10% 75,400i 7,540 1,697 9,237 Melissa Prince Lab Mgr 12 30% 45,125 13,538 3,466 17,004 Radhika Gaddipatti Fellow 12 25% 46,188 11,547 2,956 14,503 Thomas Blackwell Lab Mgr 12 25% 40,372 10,093 2,584 12,677 To be announced Res Asst III 12 100% 37,500 37,500 9,600 47,100 Kuisook Keel Technologist 12 5% 30,623 1,531 392 1,923 To be announced Res Asst II 12 100% 32,500 32,500 8,320 40,820 SUBTOTALS 189,459 43,186 232,645 CONSULTANT COSTS 0 EQUIPMENT (Itemize) 0 SUPPLIES (Itemize by category) Genotyping for Anticipation Studies 7,000 Genotyping and Sequencing for BMPR2 studies 27,000 LOH Study Supplies 400 Southern Blotting Supplies 600 Tissue Culture Supplies 15,000 Molecular Biology Supplies (disposables, plastics, glassware, pipettes, etc) 14,500 General Lab Supplies 11,000 75,500 TRAVEL 0 PATIENT CARE COSTS 0 INPATIENT OUTPATIENT 0 ALTERATIONS AND RENOVATIONS (itemize by category) None 0 OTHER EXPENSES (Itemize by category) Software Upgrades 1,000 lf000 SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD $30e,1451 0 CONSORTIUM/CONTRACTUAL DIRECT COSTS COSTS FACILITIES AND ADMINISTRATION COSTS 0 TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD (Item7a,FacePage) $309,1451 SBIR/STTR Only: FIXED FEE REQUESTED PHS398(Rev.05/01) (Page) 19 9 Form Page 4 Number pages consecutivelyat the bottom throughoutthe application. Do not usesuffixes such as 3a, 3b.