Vascular remodeling in patients with rare genetic disorders

罕见遗传性疾病患者的血管重塑

基本信息

批准号：
10929131
负责人：
Manfred Boehm
金额：
$ 182.63万
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10929131
关键词：
5&apos -Nucleotidase Acceleration Adhesions Adult Affect Aneurysm Arterial Intimas Arteries Atherosclerosis Atherosclerosis Risk in Communities Benign Biological Assay Biological Models Blood Vessels Blood flow CADASIL COVID-19 patient Calcium Candidate Disease Gene Cardiac Cell Line Cells Childhood Choristoma Chronic Chronic Disease Chronic Kidney Failure Chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature Clinical Clinical Research Coculture Techniques Collaborations Collagen Communication Complex Complication Connective Tissue Cutaneous DNA Sequence Alteration Dasatinib Data Data Analyses Defect Degos disease Dermal Diagnostic Dilatation - action Disease Disease Outcome Disease Progression Disease model Drug Screening Echocardiography Electrocardiogram Endothelial Cells Endothelium Enrollment Enzymes Etiology Evaluation Exons Extracellular Matrix Extravasation Face Fatal Outcome Fibroblasts Fibrosis Functional disorder Gene Mutation General Population Genes Genetic Grant Granulation Tissue Haplotypes Health Hepatomegaly Immune Immune system Immunologic Deficiency Syndromes In Vitro Infection Inflammation Inherited Integrins Intellectual functioning disability Intercellular adhesion molecule 1 Job&apos s Syndrome Joints LYN gene Life Liver Fibrosis Lower Extremity Lung Magnetic Resonance Imaging Manuscripts Maps Mediating Memory Loss Mesenchymal Metabolic Pathway Metabolism Microvascular Dysfunction Microvascular Permeability Migraine Modeling Molecular Multiple Abnormalities Mutation Myocardial Infarction NOTCH3 gene National Heart, Lung, and Blood Institute Natural History Nature Neonatal Onset Multisystem Inflammatory Disease Neutrophil Infiltration Non-Insulin-Dependent Diabetes Mellitus Nucleotidases Organ Outcome Oxygen Pathway interactions Patients Perinatal Perivascular Fibrosis Pharmaceutical Preparations Phase Phenotype Phosphotransferases Population Proline Proteins Proteomics Publishing Pulmonary Hypertension Purines Rare Diseases Reporting Research Research Technics Resources Risk Factors Role STAT3 gene Signal Pathway Signal Transduction Spleen Stroke Subarachnoid Hemorrhage System Systemic hypertension TBK1 gene TNF gene Technology Testing Therapeutic Time Tissues Treatment Protocols Vascular Diseases Vascular Endothelial Cell Vascular blood supply Vascular calcification Vascular remodeling Vascularization Vasculitis Xaa-Pro dipeptidase angiogenesis arterial tortuosity autoinflammatory diseases autosome biobank biomarker identification body system calcification clinical trial analysis database of Genotypes and Phenotypes disease-causing mutation effectiveness testing extracellular gain of function mutation genetic testing healing health record hypoperfusion hypoxia inducible factor 1 imaging study improved in vivo induced pluripotent stem cell inflammatory modulation inhibitor insight kinase inhibitor large datasets migration mouse model neutrophil optical imaging post-COVID-19 programs psychiatric symptom rare genetic disorder recessive genetic trait recurrent infection research clinical testing screening side effect single cell sequencing skin lesion skin ulcer slug src-Family Kinases systemic inflammatory response therapeutic target tool transcriptome sequencing treatment strategy vascular abnormality whole genome wound healing

项目摘要

Monogenetic disorders grant the unique ability to understand more complex diseases due to a single defined genetic defect. Advanced sequencing technology accelerates gene candidate discovery but progress can be slowed by the inability to distinguish between functional disease related and non-disease related mutations. We use patient-specific in vitro disease modeling systems combined with genetic tools to identify disease related mutation, to study gene related disease mechanism and to perform drug screening. Further, comprehensive clinical evaluation of patients with these conditions further help elucidate the disease mechanism and the organ systems affected. Vascular calcification is a secondary complication to diseases such as atherosclerosis, diabetes mellitus type II and chronic kidney disease but its underlying mechanism is poorly understood. ACDC is a rare disease, in which de novo vascular calcifications form in lower extremity arteries and peri-articular calcifications in the joints of affected adults and is caused by mutations in the 5'-nucleotidase Ecto (NT5E) gene encoder for CD73, an enzyme in the extracellular purine metabolic pathway. Our small non-randomized treatment protocol in seven patients with ACDC with etidronate that was completed in 2021 tested effectiveness of etidronate in attenuating the progression of lower extremity arterial/peri-articular calcification formation and improving vascular blood flow. Results showed that etidronate treatment did not significantly change these parameters over time but may have slowed the rate disease progression. Patients tolerated the medication well with no significant side effects. A manuscript summarizing this data is currently under review. Autosomal dominant hyper-IgE syndrome (AD-HIES; Jobs syndrome) is a rare immunodeficiency due to mutations in the STAT3 gene. Patients suffer from multiple life-threatening infections from childhood as well as multiple abnormalities outside the immune system, such as aberrant healing. Vascular abnormalities include arterial tortuosity and abnormal dilatation and aneurysms of medium sized arteries, potentially leading to myocardial infarction and subarachnoid hemorrhage. In a wound healing assay, we found delayed granulation tissue formation and vascularization in AD-HIES patients compared to healthy subjects. RNA-Seq analysis identified deficiencies in angiogenesis, extracellular matrix metabolism, and wound healing signaling pathways mediated by dysregulation of HIF1 signaling. Currently, we continue to look for therapeutic targets for AD-HIES patients within signaling pathways affected by deficient HIF1 signaling. Prolidase deficiency (PD) is an autosomal recessive genetic disease caused by mutations in the PEPD gene encoding the enzyme prolidase D, leading to defects in turnover of proline-containing proteins, such as collagen. PD is characterized by chronic severe skin ulcers, recurrent infections, unusual facial features, variable intellectual disability levels, and enlargement of the liver and spleen. To determine the wound healing phenotype in PD patients, we established PD patient-specific iPSC-derived endothelial cell (EC) lines and primary dermal fibroblasts lines. We plan to use single culture/co-culture systems in combination with in vitro scratch wound healing/angiogenesis assays to determine the role of vascular endothelial cells and dermal fibroblasts in PD wound healing as well as to test different kinds of treatment. Autoinflammatory diseases, such as SAVI, DADA2, NOMID, CANDLE and LYN GOF, negatively impact blood vessels, resulting in severe tissue damage as well as fatal outcomes for those affected. These diseases are caused by genetic mutations and underlying mechanisms have not been well characterized. We investigate these through clinical evaluation, genetic testing, high content screening and single cell sequencing as well as patient derived cells to investigate the disease mechanisms with in vitro/in vivo murine models. Endothelial-to-mesenchymal transition (EndMT) is a major mechanism contributing to multiple organ fibrosis. During the reporting period, we identified that spontaneously developed SAVI iEC (induced pluripotent stem cell-derived endothelial cells). EndMT contributes to perivascular fibrosis in SAVI patient lung sections. We further determined that the STING/TBK1/STAT3/Slug pathway mediates EndMT and identified potential treatments including STING, TBK1 and JAK/STAT inhibitors using a SAVI-iEC disease model. We also established an iEC/iSMC coculture in-vitro disease model for CANDLE (chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperatures), which is associated with systemic and pulmonary hypertension. This model will allow us to further explore underlying disease mechanisms and test potential therapeutic tools for CANDLE. Neutrophilic inflammation is a hallmark of many monogenic autoinflammatory diseases. The pathomechanisms that regulate extravasation of damaging immune cells into surrounding tissues are poorly understood. In the last year, we identified three unrelated patients with perinatal-onset of neutrophilic cutaneous small vessel vasculitis and systemic inflammation caused by constitutive activation of Lyn kinase. Functional studies revealed increased expression of ICAM-1 on iECs and of 2-integrins on patient neutrophils, implicated in increased neutrophil adhesion and vascular transendothelial migration (TEM). Treatment with TNF inhibition improved systemic inflammation and liver fibrosis resolved on treatment with the Src kinase inhibitor dasatinib. Our findings reveal a critical role for Lyn kinase in modulating inflammatory signals, regulating microvascular permeability and neutrophil recruitment, and in promoting hepatic fibrosis. These results were published on Nature Communications in March 2023. Small vessel diseases are conditions with narrowing of small arteries leading to an imbalance of blood supply upon demand, resulting in progressive chronic hypoperfusion with detrimental outcomes. CADASIL is caused by NOTCH3 mutations with predominant clinical features including migraines, strokes, memory loss, and multiple psychiatric symptoms. Similarly, Kholmeier-Degos (K-D) disease is a poorly understood rare vasculopathy leading to small vessel occlusions in multiple organs. Benign K-D is specifically limited to skin lesions, while systemic K-D progresses to other organs and may quickly become fatal. We developed a robust clinical research program to characterize the etiology/natural history of CADASIL and K-D through comprehensive clinical and molecular evaluations as well as to develop patient-derived disease models to better understand their pathophysiology. To compare disease progression in patients with rare diseases with similar clinical presentation to that of more common diseases, we have incorporated population data analysis from clinical trials and health records in our vascular diseases program. Large population data has recently become available through resources such as the NHLBI BioLINCC, dbGaP or UK Biobank. We are working to develop increased expertise in obtaining and analyzing these large datasets. As a result, we have analyzed data from the Atherosclerosis Risk in Communities (ARIC) study identifying risk factors for chronic diseases (PMID: 37245480; PMID: 36599719) and are currently working to integrate this analysis of risk factors with insights related to gene-disease interactions from rare disease research. We also have on-going collaborations to evaluate post-COVID19 patients with a focus on long-term cardiac and vascular sequelae. We assess cardiac and vascular health of these patients with a variety of research techniques including electrocardiograms, echocardiograms, cardiac MRIs, optical imaging studies, NIRS, etc.

由于单一定义的遗传缺陷，单遗传性疾病赋予了理解更复杂疾病的独特能力。先进的测序技术加速了候选基因的发现，但由于无法区分功能性疾病相关和非疾病相关突变，进展可能会减慢。我们利用患者特异性体外疾病建模系统结合遗传工具来识别疾病相关突变，研究基因相关疾病机制并进行药物筛选。此外，对患有这些疾病的患者进行全面的临床评估进一步有助于阐明疾病机制和受影响的器官系统。血管钙化是动脉粥样硬化、II 型糖尿病和慢性肾病等疾病的继发并发症，但其潜在机制尚不清楚。 ACDC 是一种罕见疾病，患者下肢动脉中会出现新生血管钙化，成人关节中会出现关节周围钙化，这种疾病是由 CD73（一种酶）的 5'-核苷酸酶 Ecto (NT5E) 基因编码器突变引起的。在细胞外嘌呤代谢途径中。我们于 2021 年完成的针对 7 名 ACDC 患者的小型非随机治疗方案，测试了依替膦酸在减缓下肢动脉/关节周围钙化形成进展和改善血管血流方面的有效性。结果显示，依替膦酸治疗并没有随着时间的推移显着改变这些参数，但可能减缓了疾病进展的速度。患者对药物的耐受性良好，没有明显的副作用。目前正在审查总结这些数据的手稿。常染色体显性高 IgE 综合征（AD-HIES；乔布斯综合征）是一种罕见的免疫缺陷病，由 STAT3 基因突变引起。患者从童年起就患有多种危及生命的感染，以及免疫系统之外的多种异常，例如愈合异常。血管异常包括动脉迂曲和异常扩张以及中等大小动脉的动脉瘤，可能导致心肌梗塞和蛛网膜下腔出血。在伤口愈合测定中，我们发现与健康受试者相比，AD-HIES 患者的肉芽组织形成和血管化延迟。 RNA-Seq 分析发现了 HIF1 信号失调介导的血管生成、细胞外基质代谢和伤口愈合信号通路的缺陷。目前，我们继续在受 HIF1 信号传导缺陷影响的信号通路内寻找 AD-HIES 患者的治疗靶点。脯氨酸酶缺乏症 (PD) 是一种常染色体隐性遗传病，由编码脯氨酸酶 D 的 PEPD 基因突变引起，导致含脯氨酸蛋白质（例如胶原蛋白）的周转缺陷。 PD 的特点是慢性严重皮肤溃疡、反复感染、面部特征异常、不同程度的智力障碍以及肝脏和脾脏肿大。为了确定 PD 患者的伤口愈合表型，我们建立了 PD 患者特异性 iPSC 衍生的内皮细胞 (EC) 系和原代真皮成纤维细胞系。我们计划使用单一培养/共培养系统与体外划痕伤口愈合/血管生成测定相结合，以确定血管内皮细胞和真皮成纤维细胞在PD伤口愈合中的作用，并测试不同类型的治疗方法。 SAVI、DADA2、NOMID、CANDLE 和 LYN GOF 等自身炎症性疾病会对血管产生负面影响，导致严重的组织损伤以及受影响者的致命后果。这些疾病是由基因突变引起的，其潜在机制尚未得到很好的表征。我们通过临床评估、基因测试、高内涵筛选和单细胞测序以及患者来源的细胞来研究这些疾病，以体外/体内小鼠模型研究疾病机制。内皮间质转化（EndMT）是导致多器官纤维化的主要机制。在报告期内，我们发现自发发育的 SAVI iEC（诱导性多能干细胞衍生内皮细胞）。 EndMT 导致 SAVI 患者肺切片的血管周围纤维化。我们进一步确定 STING/TBK1/STAT3/Slug 通路介导 EndMT，并使用 SAVI-iEC 疾病模型确定了包括 STING、TBK1 和 JAK/STAT 抑制剂在内的潜在治疗方法。我们还建立了 CANDLE（伴有脂肪营养不良和体温升高的慢性非典型中性粒细胞皮肤病）的 iEC/iSMC 共培养体外疾病模型，该疾病与全身性高血压和肺动脉高压相关。该模型将使我们能够进一步探索潜在的疾病机制并测试 CANDLE 的潜在治疗工具。中性粒细胞炎症是许多单基因自身炎症性疾病的标志。人们对调节破坏性免疫细胞外渗到周围组织的病理机制知之甚少。去年，我们发现了三名无关的患者，他们患有围产期发病的中性粒细胞性皮肤小血管炎和由 Lyn 激酶组成性激活引起的全身炎症。功能研究显示 iEC 上 ICAM-1 的表达增加以及患者中性粒细胞上 2-整合素的表达增加，这与中性粒细胞粘附和血管跨内皮迁移 (TEM) 的增加有关。 TNF 抑制治疗改善了全身炎症，并通过 Src 激酶抑制剂达沙替尼治疗解决了肝纤维化问题。我们的研究结果揭示了 Lyn 激酶在调节炎症信号、调节微血管通透性和中性粒细胞募集以及促进肝纤维化方面的关键作用。这些结果于 2023 年 3 月发表在《自然通讯》上。小血管疾病是小动脉狭窄导致血液供应需求不平衡的疾病，导致进行性慢性灌注不足，产生有害结果。 CADASIL 是由 NOTCH3 突变引起的，其主要临床特征包括偏头痛、中风、记忆丧失和多种精神症状。同样，Kholmeier-Degos (K-D) 病是一种人们知之甚少的罕见血管病变，会导致多个器官的小血管闭塞。良性 K-D 仅限于皮肤病变，而全身性 K-D 会进展至其他器官，并可能很快致命。我们开发了一项强大的临床研究计划，通过全面的临床和分子评估来表征 CADASIL 和 K-D 的病因/自然史，并开发源自患者的疾病模型以更好地了解其病理生理学。为了比较临床表现与常见疾病相似的罕见疾病患者的疾病进展，我们将临床试验和健康记录的人群数据分析纳入我们的血管疾病项目中。最近可通过 NHLBI BioLINCC、dbGaP 或 UK Biobank 等资源获取大量人口数据。我们正在努力提高获取和分析这些大型数据集的专业知识。因此，我们分析了社区动脉粥样硬化风险 (ARIC) 研究的数据，该研究确定了慢性疾病的风险因素（PMID：37245480；PMID：36599719），目前正在努力将风险因素的分析与与基因相关的见解结合起来。罕见疾病研究中的疾病相互作用。我们还持续合作评估新冠肺炎后患者，重点关注长期心脏和血管后遗症。我们通过各种研究技术评估这些患者的心脏和血管健康状况，包括心电图、超声心动图、心脏 MRI、光学成像研究、NIRS 等。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Development of vascular disease models to explore disease causation and pathomechanisms of rare vascular diseases.

DOI：
10.1007/s00281-022-00925-9
发表时间：
2022-05
期刊：
Seminars in immunopathology
影响因子：
9
作者：
Harper RL;Ferrante EA;Boehm M
通讯作者：
Boehm M

Diffuse atrophic papules and plaques, intermittent abdominal pain, paresthesias, and cardiac abnormalities in a 55-year-old woman.

一名 55 岁女性出现弥漫性萎缩性丘疹和斑块、间歇性腹痛、感觉异常和心脏异常。

DOI：
10.1016/j.jaad.2016.09.015
发表时间：
2016
期刊：
Journal of the American Academy of Dermatology
影响因子：
13.8
作者：
Oliver,Brittany;Boehm,Manfred;Rosing,DouglasR;Shapiro,LeeS;Dempsey,DanielT;Merkel,PeterA;Lee,Chyi-ChiaRichard;Cowen,EdwardW
通讯作者：
Cowen,EdwardW

Middle-age high normal serum sodium as a risk factor for accelerated biological aging, chronic diseases, and premature mortality.

DOI：
10.1016/j.ebiom.2022.104404
发表时间：
2023-01
期刊：
EBIOMEDICINE
影响因子：
11.1
作者：
Dmitrieva, Natalia I.;Gagarin, Alessandro;Liu, Delong;Wu, Colin O.;Boehm, Manfred
通讯作者：
Boehm, Manfred

4D Printed Cardiac Construct with Aligned Myofibers and Adjustable Curvature for Myocardial Regeneration.

DOI：
10.1021/acsami.0c17610
发表时间：
2021-03-24
期刊：
ACS APPLIED MATERIALS & INTERFACES
影响因子：
9.5
作者：
Wang, Yue;Cui, Haitao;Wang, Yancheng;Xu, Chengyao;Esworthy, Timothy J.;Hann, Sung Yun;Boehm, Manfred;Shen, Yin-Lin;Mei, Deqing;Zhang, Lijie Grace
通讯作者：
Zhang, Lijie Grace

The benefits and risks of stem cell technology.

干细胞技术的好处和风险。