Advancing Small Molecule Read Through Compounds to Prevent and/or Treat Heritable Pulmonary Artery Hypertension

推进小分子化合物读取以预防和/或治疗遗传性肺动脉高血压

基本信息

批准号：
10011012
负责人：
VIRENDER K REHAN
金额：
$ 25.21万
依托单位：
ADVENT THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-15 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10011012
关键词：
Activities of Daily Living Acute Address Aftercare Amino Acids Anatomy Antibiotics BMPR2 gene Biochemical Biological Assay Blood Blood Cells Bone Morphogenetic Protein Receptor Gene Cardiovascular Diseases Catheterization Cell membrane Cell surface Cells Cellular Assay Cessation of life Characteristics Childhood Chronic Chronic Toxicity Tests Collaborations DNA Sequence Alteration Data Development Disease Dose Endothelial Cells Endothelium Exhibits Functional disorder Gene Targeting Genes Heart Heart failure Hereditary Disease Heritability Human In Vitro Individual Lead Legal patent Length Life Lung Medical Messenger RNA Methods Mus Mutation Names Neonatal Nonsense Mutation Orphan Drugs Pathology Pathway interactions Patients Permeability Pharmaceutical Preparations Phosphorylation Positioning Attribute Prevalence Production Proteins Publishing Pulmonary Hypertension Pulmonary artery structure Pulmonary vessels Rare Diseases Research Personnel Ribosomes Rodent Series Signal Pathway Signal Transduction Site Smooth Muscle Myocytes Solubility Terminator Codon Testing Therapeutic Tissues Toxic effect Transcript Treatment Effectiveness Treatment Efficacy Universities Up-Regulation base bone morphogenetic protein receptors cadherin 5 clinically relevant dalton effective therapy efficacy testing experimental study hemodynamics high throughput screening improved in vitro Assay in vivo mRNA Expression metabolic profile mortality mouse model multidisciplinary mutant novel therapeutics patient population premature pressure prevent protein expression pulmonary arterial hypertension receptor receptor expression reflectance confocal microscopy small molecule small molecule libraries

项目摘要

Advent Therapeutics is a company that develops new therapies for neonatal, pediatric, and Orphan Drug “niche” markets to address serious unmet medical needs. Our team proposes early stage development of a new treatment for hereditable pulmonary arterial hypertension (hPAH), a rare, hereditary disorder that involves progressive elevation of pulmonary arterial pressures, right heart failure, and death. There are no effective treatments. hPAH is often caused by premature termination codons (PTCs) in the mRNA encoded by nonsense mutations in the type II bone morphogenetic protein receptor (BMPR2) gene. Certain antibiotics possess the capacity to induce “readthrough” of PTCs and thus produce a full-length protein, but are unacceptably toxic. To obviate this toxicity, we first performed a high-throughput screen on a 70,000-compound small molecule library, using a readthrough assay that indicated hits. We derivatized and patented hits, achieved in vitro, ex vivo, and in vivo proof-of-concept using PTCs in disease-causing genes other than BMPR2, and published these results. We next selected a molecule named GJ103 from among the derivatized compounds for further development, based on readthrough efficiency and favorable physical and solubility characteristics, and began testing in hPAH, using blood outgrowth endothelial cells obtained from patients with hPAH that contained disease-relevant PTCs in BMPR2. GJ103 treatment induced significant expression of BMPR2 protein. Further testing with mouse cells corroborated the results with human cells. Next, we collaborated with the Morell lab (University of Cambridge, UK) to test GJ103 in mice they created that develop hPAH by virtue of one of two clinically-relevant nonsense mutations in Bmpr2 (Bmpr2+/R899X and Bmpr2+/R584X mice). After performing preliminary dose-finding studies, GJ103 induced up to half of wild-type BMPR2 protein levels in vivo. Preliminary acute and chronic toxicity testing failed to show evidence of toxicity. BMPR2 protein induced by GJ103 demonstrated its functional effects in an LPS permeability assay in vitro and in vivo, and confocal microscopy studies indicated the protein appeared to traffic properly to the cell surface of endothelial and smooth muscle cells. Downstream upregulation of BMP signaling intermediaries (SMADs and phosphoSMADs) and BMP pathway gene targets (Id1, Id2, and VE- cadherin) indicated functional activation of BMP signaling. Here we propose to test the critical question for using GJ103 to treat hPAH: can GJ103-induced expression of BMPR2 prevent or slow development of hPAH in Bmpr2+/R899X or Bmpr2+/R584X mice if treatment is initiated early enough? In Aim 1, we will collaborate with the Soban lab (UCLA) to test whether GJ103 can induce expression of BMPR2 sufficient to slow or halt development of hPAH in mice. Dependent variables include testing right heart hemodynamics and remodeling using echo and direct catheterization in Bmpr2+/R899X and Bmpr2+/R584X mice treated with GJ103. Aim 2 proposes concomitant expression analyses with the Rehan lab (LA BioMed) of BMPR2 protein, which should be elevated if GJ103 is successful. Our studies might represent the first hope for individuals suffering from hPAH.

Advent Therapeutics是一家为新生儿，儿科和孤儿开发新疗法的公司 “利基市场”市场以满足严重的未满足医疗需求。我们的团队提案的早期舞台发展治疗可遗传的肺动脉高压（HPAH），这是一种罕见的遗传性疾病，涉及肺动脉压力，右心力衰竭和死亡的进行性升高。没有有效治疗。 HPAH通常是由胡说八道编码的mRNA中的过早终止密码子（PTC）引起的 II型骨形态发生蛋白受体（BMPR2）基因中的突变。某些抗生素具有诱导PTC“读取”并因此产生全长蛋白的能力，但具有不可接受的毒性。到消除这种毒性，我们首先在70,000个混合小分子库上进行了高通量屏幕，使用指示命中的读取测定法。我们衍生和获得专利的命中，在体外实现，离体和在BMPR2以外的其他引起疾病基因中使用PTC的体内概念验看，并发表了这些结果。接下来，我们从衍生化合物中选择了一个名为GJ103的分子，以进行进一步开发，基于读取效率和有利的身体和溶解度特征，并开始在HPAH进行测试，使用从含有与疾病相关的PTC的HPAH患者获得的血液生长的内皮细胞在BMPR2中。 GJ103处理诱导BMPR2蛋白的显着表达。用小鼠细胞进行进一步测试通过人类细胞证实了结果。接下来，我们与莫雷尔实验室（剑桥大学，英国）在小鼠中测试GJ103，他们创建了通过两个临床上的胡说八道之一来发展HPAH BMPR2（BMPR2+/R899X和BMPR2+/R584X小鼠）中的突变。在进行初步剂量调查研究之后， GJ103在体内诱导了多达一半的野生型BMPR2蛋白水平。初步急性和慢性毒性测试未能显示毒性的证据。 GJ103诱导的BMPR2蛋白证明了其在A中的功能作用 LPS的体外和体内评估，共聚焦显微镜研究表明蛋白质似乎是正确地流动到内皮和平滑肌细胞的细胞表面。 BMP的下游上调信号中介机构（SMADS和Phosphosmads）和BMP途径基因靶标（ID1，ID2和VE- Cadherin）表示BMP信号传导的功能激活。在这里，我们建议测试使用的关键问题 GJ103治疗HPAH：GJ103诱导的BMPR2的表达可以防止HPAH在 BMPR2+/R899X或BMPR2+/R584X小鼠如果较早开始治疗？在AIM 1中，我们将与 SOBAN LAB（UCLA）测试GJ103是否可以诱导BMPR2的表达足以缓慢或停止小鼠HPAH的发展。因变量包括测试右心血位动力学和重塑在BMPR2+/R899X和BMPR2+/R584X小鼠中使用ECHO和直接导管插入，用GJ103处理。目标2提案 BMPR2蛋白的Rehan Lab（LA Biomed）伴随表达分析，应升高如果GJ103成功。我们的研究可能代表了患有HPAH的人的第一个希望。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Maternal food restriction-induced intrauterine growth restriction in a rat model leads to sex-specific adipogenic programming.

DOI：
10.1096/fj.202000985rr
发表时间：
2020-12
期刊：
FASEB journal : official publication of the Federation of American Societies for Experimental Biology
影响因子：
0
作者：
Sreekantha S;Wang Y;Sakurai R;Liu J;Rehan VK
通讯作者：
Rehan VK

Perinatal exposure to nicotine alters spermatozoal DNA methylation near genes controlling nicotine action.