"Exploration of Human Parathyroid Cellular Organization and Function"

项目摘要 甲状旁腺功能低下（HP）是甲状腺恶性肿瘤手术的主要毒性反应。甲状旁腺（PT）是 内分泌器官对维持钙稳态至关重要。HP最常见的原因是医源性的- 发生在切除PT腺体后，在前和中央颈室的解剖过程中， 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发展为HP 甲状腺切除术，并且在美国患有HP的成年人总数约为77，000人。有些患者 不能维持稳定的生理水平的Ca 2+，并可能遭受严重的后果，包括肌肉 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PT干/祖细胞的鉴定和特性 对于开发有效的HP细胞治疗方法至关重要。克里希纳斯瓦米实验室 开发了一种新的计算工具，PHATE（势的热扩散的亲和力为基础的转变 嵌入），其能够捕获内在数据几何形状并对scRNA-seq数据进行降噪，使得局部和非局部数据都能够被捕获。 和全球结构是显而易见的，而无需对数据的形式强加假设。PHATE是 在分析由动态过程生成的微分数据时， 数据点之间的连续变化，因此是一个理想的工具，用它来解剖细胞 复杂组织的组成，如PT腺。我们假设PT腺体的维持 依赖于位于支持性小生境内的干/祖细胞区室。我们建议定义这些 使用scRNA-seq，PHATE和功能性的细胞组分和PT组织的维持机制 通过两个具体目标进行研究。首先，我们将获得初级PT腺体并进行scRNA-Seq， 然后将与PHATE分析，以重建细胞的组织和维护机制， 人PT腺。在第二部分中，我们将确定特定细胞的PT腺体内的功能 由PHATE确定的人口。PT细胞亚群的长期体内功能将通过 移植到免疫缺陷小鼠中。这些研究将有助于阐明 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， 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

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.