"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.

项目摘要 甲状腺功能减退症（HP）是手术对甲状腺恶性肿瘤的主要毒性。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的最常见原因是医源性 - 在解剖过程中切除PT腺后发生后发生的前后颈部室室的发生后发生的 甲状腺（或甲状旁腺）癌症的治疗。据估计，有5-7％的患者随后出现HP 甲状腺切除术，美国HP成年人的总数约为77,000。有些患者是 无法维持稳定的Ca2+生理水平，并且可能遭受严重的后果，包括肌肉 抽筋，四分和抽搐。克劳斯实验室正在开发区分方法 自体细胞进入功能性PT细胞。人类PT茎/祖细胞的识别和表征 对于开发HP细胞疗法的有效方法至关重要。 Krishnaswamy实验室有 开发了一种新型的计算工具Phate（基于亲和力的过渡的热水扩散的潜力 嵌入），它能够捕获固有的数据几何并denoise scrna-seq数据，以便两个局部 并且全球结构是显而易见的，而没有对数据形式施加假设。 Phate是 在分析由有动态过程产生的分化数据方面特别有力的 数据点之间的连续变异性，因此是剖析细胞的理想工具 复杂组织（例如Pt腺）的组成。我们假设PT腺的维护 依靠位于支撑壁球中的茎/祖细胞室。我们建议定义这些 使用SCRNA-SEQ，PHATE和功能 通过两个具体目标进行研究。首先，我们将获得主要的pt腺并执行scrna-seq， 然后将用PHATE分析以重建细胞组织和维护机制 人Pt腺。在第二个中，我们将确定特定细胞的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由 移植到免疫缺陷的小鼠中。提出的研究将阐明组成和细胞 不同PT隔室的功能，并将增强我们对稳态机制的理解 维持正常PT腺的功能性上皮室。获得的见解将增强 了解成人PT腺体维持和再生的基本机制，必需的临床前 了解新型细胞疗法以恢复PT功能的知识 甲状腺功能减退症。