Single-Cell Transcriptome & Effect of Immune Checkpoint Therapy on Kaposi Sarcoma

单细胞转录组及免疫检查点治疗对卡波西肉瘤的影响 卡波西肉瘤(KS)是一种由卡波西-疱疹病毒(KS-γ-HerpresVirus，KSHV)感染引起的恶性肿瘤。 免疫抑制的个体，以及一种主要发生在撒哈拉以南非洲男性和 南地中海国家。肿瘤被认为起源于淋巴管内皮细胞，但事实并非如此。 病理复杂，混合有血管样裂隙、梭形细胞和浸润性炎症细胞。 KSHV在一组具有裂解或潜伏基因表达谱的肿瘤细胞中被发现。由于这种复杂性， 关于这种恶性肿瘤的基因组和转录特征的信息非常有限。这个 单细胞(Sc)转录组学(rna-seq)的发展为识别关键基因提供了独特的机会。 KS发展的基础特征。 KS的治疗包括局部治疗、化疗、血管生成或蛋白酶体抑制剂，或者 免疫疗法，如免疫调节药物(IMids)或免疫检查点抑制剂。最新发现 认为KS对程序性死亡-1(PD-1)途径的抑制剂的反应大约是在20-20分钟内完成的。 30%的个体，但很少会引起严重的炎症性KSHV相关疾病。当前项目 将利用我们目前在HIV相关KS中进行的nivolumab试验的一个扩展分支。我们将选择 尼伏鲁单抗治疗2个月前和治疗后2个月的样本分别来自5名反应最好的患者和5名 患有进展性疾病或对治疗无效的个人。当前项目的一个目标是确定 使用scRNA-seq的前治疗特征，预测PD-KS治疗的反应或进展。 1抑制剂，尼伏卢单抗。这个项目的第二个目标是确定细胞组成和基因的变化 随着nivolumab在肿瘤细胞和肿瘤微环境中的表达。目标如下： 目的1.研究PD-1检查点抑制剂治疗对KS肿瘤淋巴组织浸润的影响。 我们将使用scRNA-seq来计数B，CD4+和CD8+淋巴细胞，自然杀伤细胞，树突状细胞和浆细胞， 尼伏单抗治疗前后样本中的巨噬细胞、间充质细胞、内皮细胞和其他细胞。 目的2.检测PD-1检查点抑制剂治疗对细胞和病毒基因表达的影响。 我们将使用scRNA-seq来量化KSHV裂解基因和潜伏基因的表达水平，以及 血管生成、增殖、细胞凋亡和代谢途径中的细胞基因。

Abstract – Single-Cell Transcriptome & Effect of Immune Checkpoint Therapy on Kaposi Sarcoma Kaposi Sarcoma (KS) is a malignancy resulting from KS γ-herpresvirus (KSHV) infection occurring in immunosuppressed individuals, as well as a disease occurring primarily in males in subSaharan Africa and southern Mediterranean countries. The tumor is thought to arise from lymphatic endothelial cells, but it is pathologically complex with an admixture of vascular-like slits, spindle cells, and infiltrating inflammatory cells. KSHV is found in a subset of tumor cells with either lytic or latent gene expression profiles. Due to this complexity, there is very limited information about genomic and transcriptomic characteristics of this malignancy. The development of single cell (sc) transcriptomics (RNA-seq) provides a unique opportunity to identify key characteristics that underlie the development of KS. Treatment of KS involves local therapies, chemotherapy, angiogenesis or proteasome inhibitors, or immunotherapies, such as immunomodulatory drugs (IMiDs) or immune checkpoint inhibitors. Recent findings have suggested that KS responds to inhibitors of the programmed death-1(PD-1) pathway in approximately 20- 30% of individuals, but may rarely cause severe inflammatory KSHV-associated disorders. The current project will take advantage of an expansion arm of our current trial of nivolumab in HIV-associated KS. We will select samples before and after 2 months of nivolumab treatment from the 5 individuals with the best response and 5 individuals with progressive disease or no response to treatment. One goal of the current project is to identify pre-treatment characteristics, using scRNA-seq, that predict response or progression of KS treatment with PD- 1 inhibitor, nivolumab. A second goal of this project is to determine what changes of cell composition and gene expression occur with nivolumab in tumor cells and the tumor microenvironment. The aims are as follows: Aim 1. Determine the effect of PD-1 checkpoint inhibitor therapy on lymphoid infiltration into KS tumors. We will use scRNA-seq to enumerate B, CD4+ and CD8+ lymphocytes, natural killer, dendritic, and plasma cells, macrophages, mesenchymal, endothelial, and other cells in samples before and after nivolumab treatment. Aim 2. Determine the effect of PD-1 checkpoint inhibitor therapy on cellular and viral gene expression. We will use scRNA-seq to quantify levels of expression of KSHV lytic and latent genes, as well as expression of cellular genes in angiogenesis, proliferation, apoptosis, and metabolic pathways.