"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.

项目总结 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)是 对维持钙平衡至关重要的内分泌器官。幽门螺杆菌最常见的原因是医源性- 发生在颈椎前间隙和中央间隙清扫术中切除甲状旁腺后 甲状腺(或甲状旁腺)癌的治疗。据估计，5-7%的患者会在以下情况下患上幽门螺杆菌 在美国，成年幽门螺杆菌患者总数约为77,000人。有些病人是 无法维持稳定的生理钙离子水平，并可能遭受包括肌肉在内的严重后果 抽筋、抽筋和抽搐。克劳斯实验室正在开发区分 将自体细胞转化为具有功能的PT细胞。人PT干/祖细胞的鉴定和鉴定 对于开发有效的幽门螺杆菌细胞疗法至关重要。Krishnaswamy实验室已经 开发了一种新的计算工具PHATE(基于亲和力的转变的热扩散势 嵌入)，它能够捕获固有的数据几何结构并对scRNA-seq数据进行去噪，从而使本地 在不对数据形式强加假设的情况下，全球结构显而易见。PHATE是 在分析由动态过程生成的差异化数据方面尤其强大 数据点之间的连续可变性，因此是解剖细胞的理想工具 复杂组织的组成，如甲状旁腺。我们假设甲状旁腺的维持 依赖于位于支持性壁龛内的干细胞/祖细胞隔间。我们建议将这些定义 利用scRNA-seq、PHATE和Function对PT组织的细胞成分和维持机制的研究 通过两个具体目标进行研究。首先，我们将获得原代PT腺体，并进行scRNA-Seq 然后用PHATE分析以重建细胞组织和维护机制 人甲状旁腺。在第二个实验中，我们将确定特定细胞的PT腺体内的功能 由PHATE确定的种群。将通过以下方法评估PT细胞亚群的长期体内功能 移植到免疫缺陷小鼠体内。所提出的研究将有助于揭示其组成和细胞 不同的PT区的功能，将加深我们对体内平衡机制的理解 维持正常甲状旁腺上皮区的功能。所获得的洞察力将增强 了解成人甲状旁腺维持和再生的基本机制 对发展新的细胞疗法以恢复患者PT功能的认识 甲状旁腺功能减退症。