Integrative Approaches to Decipher Genetic Determinants of Disease Penetrance in Prokineticin 2 Pathway Related Human Reproductive Disorders

破译原动力蛋白 2 通路相关人类生殖疾病疾病外显率遗传决定因素的综合方法

• 批准号: 10409787
• 负责人: Ravikumar Balasubramanian
• 金额: $ 33.31万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-13 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10409787
• 关键词: Address; Alleles; Amenorrhea; Anosmia; Biological; Biological Assay; Biology; Candidate Disease Gene; Chemotaxis; Circadian Rhythms; Code; Collaborations; Copy Number Polymorphism; Data Set; Defect; Development; Disease; Dissection; Dominant-Negative Mutation; Eating; Etiology; Exhibits; Fertility; G Protein-Coupled Receptor Signaling; GNRH1 gene; Genes; Genetic; Genetic Determinism; Genetic Predisposition to Disease; Genetic study; Genome; Genotype; Goals; Gonadotropin Hormone Releasing Hormone; Harvest; Heterogeneity; Hospitals; Human; Hypothalamic structure; In Vitro; Individual; Infertility; Kallmann Syndrome; Missense Mutation; Mission; Molecular; Mus; Mutation; Neurons; Neurosecretory Systems; Pain; Pathogenicity; Pathway interactions; Patients; Penetrance; Phenotype; Physiological; Physiological Processes; Population; Population Control; Regulator Genes; Reproduction; Resources; Role; SNP genotyping; Severities; Sexual Maturation; Signal Transduction; Spermatocytes; United States National Institutes of Health; Variant; angiogenesis; biobank; blood glucose regulation; cohort; combinatorial; comparative; corpus luteum; differential expression; energy balance; exome sequencing; experimental study; gene regulatory network; genetic variant; glucose metabolism; hormone deficiency; individual variation; insight; migration; mouse model; mutant; neutrophil; novel; personalized medicine; phenome; phenomics; phenotypic data; precision genomic medicine; receptor; reproductive; reproductive system disorder; transcriptomics

A major challenge in accomplishing genomic precision medicine is the observation that genotype does not always predict phenotype. Incomplete penetrance and variable expressivity represent two examples of this phenotypic variability. This complexity is well-illustrated by human mutations in PROK2 (encoding prokineticin 2) and PROKR2 (prokineticin receptor 2), that cause human Isolated GnRH (gonadotropin-releasing hormone) Deficiency (IGD). Prokineticin 2 signaling is a master regulator of mammalian reproduction and is associated with neurodevelopmental and neuroendocrine forms of IGD in humans (Kallmann syndrome, and normosmic IGD, respectively). In addition to this physiologic complexity, PROK2/R2 mutations are predominantly missense mutations that exhibit either reduced penetrance and/or variable expressivity. The genetic basis of this variability is not fully understood but allelic heterogeneity and modifier genes have been postulated to account for this variability. In Specific Aim 1, allelic heterogeneity of human PROKR2 mutations will be examined in vitro, using PROK2 signaling assays and allele specific expression studies. Whole exome sequencing and SNP genotyping will identify modifier genes that contribute to disease penetrance and expressivity. PROK2 is also implicated in diverse physiologic processes such as glucose metabolism, energy balance, food intake, circadian rhythms, pain modulation, neutrophil chemotaxis and angiogenesis. The human relevance of these pleotropic functions of PROK2 is yet to be established. In Specific Aim 2, phenomic dissection of humans with PROK2/R2 mutations will be undertaken to define the pleotropic phenotypes relating to PROK2/R2 mutations in humans. IGD patients as well as control individuals in a tertiary hospital biobank population will be studied. Reproductive phenotypes and non-reproductive phenotypes will be charted. Targeted phenotyping will include assessment of glucose homeostasis and pain modulation in both groups. A phenome-wide association study will be conducted in the biobank subjects harboring PROK2/R2 mutations. In Specific Aim 3, using a novel PROKR2-cre mouse model, transcriptomic studies of PROKR2 and GnRH neurons will be done at various developmental timepoints using temporal inhibition of PROKR2 using chemogenetics. Sexual maturation and fertility of these mice after temporal PROKR2 inhibition will also be examined. The top genes that are differentially expressed in the murine transcriptomic experiments will be translationally validated by assessing whether these genes emerge as modifier genes in the human whole exome sequencing studies. The studies in this proposal have the potential to unravel novel genetic mechanisms of disease penetrance relating to the PROK2 pathway mutations, uncover potential pleotropic human phenotypes associated with mutations in this pathway and help deliver personalized medicine to humans with reproductive disorders and infertility.

实现基因组精准医学的一个主要挑战是观察到基因型并不 总是预测表型。不完全外显率和可变表达率代表了这方面的两个例子 表型变异。这种复杂性在 PROK2（编码前动力蛋白）的人类突变中得到了很好的说明。 2) 和 PROKR2（前动力蛋白受体 2），引起人类分离 GnRH（促性腺激素释放激素） 缺乏症（IGD）。 Prokineticin 2 信号传导是哺乳动物繁殖的主要调节因子，与 人类 IGD 的神经发育和神经内分泌形式（卡尔曼综合征和正常渗透压 分别为 IGD）。除了这种生理复杂性之外，PROK2/R2 突变主要是错义的 表现出外显率降低和/或表达性可变的突变。这种变异的遗传基础 尚未完全了解，但已假设等位基因异质性和修饰基因可以解释这一点 可变性。在具体目标 1 中，将使用体外方法检查人类 PROKR2 突变的等位基因异质性 PROK2 信号传导测定和等位基因特异性表达研究。全外显子组测序和 SNP 基因分型 将识别有助于疾病外显率和表达性的修饰基因。 PROK2 也涉及 不同的生理过程，如葡萄糖代谢、能量平衡、食物摄入、昼夜节律、疼痛 调节、中性粒细胞趋化性和血管生成。这些多效性功能的人类相关性 PROK2 尚未建立。在特定目标 2 中，对具有 PROK2/R2 突变的人类进行表组解剖 将致力于定义与人类 PROK2/R2 突变相关的多效性表型。 IGD患者 以及三级医院生物库人群中的对照个体也将被研究。生殖表型 和非生殖表型将被绘制成图表。目标表型分析将包括葡萄糖评估 两组的体内平衡和疼痛调节。一项全表型关联研究将在 生物库受试者携带 PROK2/R2 突变。在 Specific Aim 3 中，使用新型 PROKR2-cre 小鼠模型， PROKR2 和 GnRH 神经元的转录组学研究将在不同的发育时间点使用 使用化学遗传学暂时抑制 PROKR2。这些小鼠的性成熟和生育能力 还将检查 PROKR2 抑制。小鼠中差异表达最高的基因 转录组实验将通过评估这些基因是否出现来进行翻译验证 人类全外显子组测序研究中的修饰基因。本提案中的研究具有潜力 揭示与 PROK2 通路突变相关的疾病外显率的新遗传机制，揭示 与该途径突变相关的潜在多效性人类表型，并有助于提供个性化 用于患有生殖障碍和不孕症的人类的药物。

项目成果

Hypothalamic Ceramides and the Ovarian Sympathetic System: At the Crossroads of Obesity and Puberty.

• DOI: 10.1016/j.cmet.2020.11.012
• 发表时间: 2021-01-05
• 期刊: Cell metabolism
• 影响因子: 29
• 作者: Stamou MI;Balasubramanian R
• 通讯作者: Balasubramanian R

Low Perinatal Androgens Predict Recalled Childhood Gender Nonconformity in Men.

围产期雄激素水平低预示着男性童年时期性别不合格的回忆。

• DOI: 10.1177/09567976211036075
• 发表时间: 2022
• 期刊: Psychological science
• 影响因子: 8.2
• 作者: Shirazi,TaliaN;Self,Heather;Rosenfield,KevinA;Dawood,Khytam;Welling,LisaLM;Cárdenas,Rodrigo;Bailey,JMichael;Balasubramanian,Ravikumar;Delaney,Angela;Breedlove,SMarc;Puts,DavidA
• 通讯作者: Puts,DavidA

Timing of peripubertal steroid exposure predicts visuospatial cognition in men: Evidence from three samples.

• DOI: 10.1016/j.yhbeh.2020.104712
• 发表时间: 2020-05
• 期刊: Hormones and behavior
• 影响因子: 3.5
• 作者: Shirazi TN;Self H;Cantor J;Dawood K;Cárdenas R;Rosenfield K;Ortiz T;Carré J;McDaniel MA;Blanchard R;Balasubramanian R;Delaney A;Crowley W;Breedlove SM;Puts D
• 通讯作者: Puts D

Behind the scenes: epigenetic mechanisms rule the roost in pubertal timing.

• DOI: 10.1016/s2213-8587(23)00167-5
• 发表时间: 2023-06
• 期刊: The lancet. Diabetes & endocrinology
• 作者: Ravikumar Balasubramanian

Evidence that perinatal ovarian hormones promote women's sexual attraction to men.

有证据表明，围产期卵巢激素会促进女性对男性的性吸引力。

• DOI: 10.1016/j.psyneuen.2021.105431
• 发表时间: 2021-09-27
• 期刊: Psychoneuroendocrinology
• 影响因子: 3.7
• 作者: Shirazi TN;Self H;Dawood K;Welling LLM;Cárdenas R;Rosenfield KA;Bailey JM;Balasubramanian R;Delaney A;Breedlove SM;Puts DA
• 通讯作者: Puts DA