"Exploration of Human Parathyroid Cellular Organization and Function"

《人类甲状旁腺细胞组织与功能的探索》

• 批准号: 10044664
• 负责人: Diane S Krause
• 金额: $ 43.07万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-20 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10044664
• 关键词: Adult; Affinity; Allogenic; Anterior; Autologous; Autologous Transplantation; Benign; Cadaver; Calcium; Calcium-Sensing Receptors; Cell Compartmentation; Cell Therapy; Cell physiology; Cells; Chief Cell; Complex; Convulsions; Data; Development; Diet; Diffusion; Dissection; Endocrine Glands; Endothelial Cells; Engraftment; Epithelial; Epithelial Cells; Epithelium; Excision; Expression Profiling; Eye; Gene Expression; Gene Expression Profile; Geometry; Heterogeneity; Histologic; Homeostasis; Hormone secretion; Human; Hypoparathyroidism; Iatrogenesis; Immunodeficient Mouse; Immunofluorescence Immunologic; Intestines; Kidney; Knowledge; Laboratories; Light; Maintenance; Malignant Neoplasms; Malignant neoplasm of thyroid; Measures; Methods; Mission; Modeling; Molecular; Muscle; Muscle Cramp; Natural regeneration; Neck; Operative Surgical Procedures; Oxyphil Cells; PTH gene; Parathyroid Adenoma; Parathyroid gland; Patients; Physiological; Population; Postoperative Period; Process; Public Health; Research; Sampling; Secondary to; Serum; Site; Stromal Cells; Structure; Surface; Surgeon; Technology; Tetany; Thyroid Gland; Thyroidectomy; Tissues; Toxic effect; Transplantation; United States National Institutes of Health; Vitamin D; absorption; arm; base; bone; burden of illness; cancer surgery; candidate marker; cell type; computerized tools; denoising; disability; human disease; in vivo; induced pluripotent stem cell; insight; malignant parathyroid gland tumor; novel; post-transplant; pre-clinical; progenitor; prospective; reconstitution; regenerative therapy; repaired; response; single-cell RNA sequencing; stem; stem cells; tool

PROJECT SUMMARY Hypoparathyroidism (HP) is a major toxicity of surgery for thyroid malignancy. Parathyroid (PT) glands are endocrine organs critical for maintaining calcium homeostasis. The most common cause of HP is iatrogenic - occurring after removal of the PT glands during dissection of the anterior and central neck compartment for the treatment of thyroid (or parathyroid) cancer. It is estimated that 5-7% of patients develop HP following thyroidectomy, and that the total number of adults with HP in the US is about 77,000. Some patients are unable to maintain stable physiological levels of Ca2+ and can suffer severe consequences including muscle cramping, tetany, and convulsions. The Krause laboratory is developing approaches for differentiation of autologous cells into functional PT cells. Identification and characterization of human PT stem/progenitor cells is critical for developing effective approaches for cell therapies for HP. The Krishnaswamy laboratory has developed a novel computational tool, PHATE (Potential of Heat-diffusion for Affinity-based Transition Embedding), which is able to capture intrinsic data geometry and denoise scRNA-seq data so that both local and global structures are apparent without imposing assumptions on the form of the data. PHATE is particularly powerful in analyzing differentiation data generated by dynamic processes where there is continuous variability between data points and thus is an ideal tool with which to dissect the cellular composition of complex tissues such as the PT gland. We hypothesize that the maintenance of the PT glands relies on stem/progenitor cell compartments that reside within supportive niches. We propose to define these cellular components and the maintenance mechanisms of PT tissue using scRNA-seq, PHATE, and functional studies through two specific aims. In the first, we will obtain primary PT glands and perform scRNA-Seq, which will then be analyzed with PHATE to reconstruct the cellular organization and the maintenance mechanisms of human PT gland. In the second, we will determine the functionality within the PT glands of specific cell populations determined by PHATE. Longterm in vivo function of PT cell subpopulations will be assessed by transplantation into immunodeficient mice. The studies proposed will shed light on the composition and cellular functions of different PT compartments, and will enhance our understanding of the homeostatic mechanisms that maintain the functional epithelial compartments of normal PT glands. The insight obtained will enhance understanding of the basic mechanisms of adult PT gland maintenance and regeneration, essential preclinical knowledge for the development of novel cellular therapies to restore PT function to patients with hypoparathyroidism.

项目概要 甲状旁腺功能减退症（HP）是甲状腺恶性肿瘤手术的主要毒性反应。甲状旁腺 (PT) 腺体是 内分泌器官对于维持钙稳态至关重要。 HP 最常见的原因是医源性 - 在解剖前部和中央颈室期间去除 PT 腺体后发生 治疗甲状腺（或甲状旁腺）癌。据估计，5-7% 的患者在以下情况下会出现 HP 甲状腺切除术，美国患有 HP 的成年人总数约为 77,000 人。有些患者是 无法维持稳定的 Ca2+ 生理水平，可能会遭受包括肌肉在内的严重后果 痉挛、手足抽搐和抽搐。克劳斯实验室正在开发区分 自体细胞转化为功能性PT细胞。人类 PT 干/祖细胞的鉴定和表征 对于开发 HP 细胞疗法的有效方法至关重要。克里希纳斯瓦米实验室 开发了一种新型计算工具 PHATE（基于亲和力的热扩散潜力 Embedding），能够捕获内在数据几何并对 scRNA-seq 数据进行去噪，以便本地 全局结构是显而易见的，无需对数据形式强加假设。 PHATE 是 在分析动态过程生成的差异数据方面特别强大 数据点之间的连续变化，因此是剖析细胞的理想工具 复杂组织的组成，如 PT 腺体。我们假设 PT 腺体的维持 依赖于位于支持性生态位内的干/祖细胞区室。我们建议定义这些 使用 scRNA-seq、PHATE 和功能性分析 PT 组织的细胞成分和维持机制 通过两个具体目标进行研究。首先，我们将获得原发性 PT 腺体并进行 scRNA-Seq，这 然后用 PHATE 进行分析，以重建细胞组织和维持机制 人类 PT 腺。第二步，我们将确定特定细胞 PT 腺内的功能 由 PHATE 确定的种群。 PT细胞亚群的长期体内功能将通过以下方式进行评估 移植到免疫缺陷小鼠体内。拟议的研究将揭示其组成和细胞 不同 PT 室的功能，并将增强我们对稳态机制的理解 维持正常 PT 腺体的功能性上皮区室。获得的洞察力将增强 了解成人 PT 腺体维持和再生的基本机制，临床前必不可少 开发新型细胞疗法以恢复患者 PT 功能的知识 甲状旁腺功能减退症。