Dissecting cell composition and drug sensitivity in human adenoid cystic carcinomas (ACCs).

摘要腺样囊性癌是最常见的生物学侵袭性癌之一。 涎腺上皮性恶性肿瘤，目前尚无临床批准的治疗方法。ACCS 具有较高的神经周围浸润和远处转移的趋势，并主要 对传统的化疗和放射治疗方法无效。ACC也由一种独特的生物学定义 特征：恶性组织中两种不同表型的癌细胞亚群共存 (导管样vs.肌上皮样)具有截然不同的基因表达谱和功能 属性。这项研究将利用我们实验室最近确定的一系列表面标记， 首次允许对这两种细胞类型进行差别化纯化、分子研究和功能分析。 本研究正式检验的假设是：a)这两种细胞类型以动态和层次化的方式共存 平衡，一种细胞类型充当另一种细胞类型的祖细胞；以及b)生化途径 调节一种细胞类型到另一种细胞类型的转换可以被用来开发新的治疗方法 对ACCS的攻击。该项目设想了两个具体目标：1)阐明发展关系 在ACCs的两个细胞成分之间：我们将使用荧光激活细胞分选(FACS)来 从人AC细胞中提纯两个亚群的癌细胞，然后在体内采用前瞻性的方法 异种移植实验和单细胞rna-seq技术，以测试两种细胞类型：a) 在患者来源的异种移植(PDX)中显示出明显的系统性差异 来自独立患者的队列；b)是等级相关的(即，两者是否可以通过整形相互转化为 彼此或其中一个是否以单向方式充当另一个的祖先)；c)包含 额外的，以前未被识别的细胞亚群，它们本身的特征是不同的分子和 功能特性；2)检测新型药物组合的体内外治疗效果 针对ACCs，旨在针对显示出影响差异表示的生化途径 和优先存活的两个细胞亚型：我们将处理3D有机化合物和PDX株显示 用序贯药物方案概括原代AC细胞的双重细胞组成，设计在第一步， 诱导肌上皮样细胞分化为导管样细胞，并在第二步选择性地 消除导管样细胞；这种新的药物方案将被测试其能力：a)导致 恶性组织的细胞成分(导管/肌上皮细胞比率)；b)缩小肿瘤大小和/或生长动力学； C)防止体内转移扩散(使用非侵入性生物发光记者)。意义重大。这 这项研究将阐明在人类ACCs中发现的两种细胞类型的分子和功能特性，填补了 在唾液肿瘤生物学方面的主要科学空白，并解决唾液腺肿瘤学中未得到满足的临床需求： 用于ACCs药物治疗的新药物靶点的确定。

Adenoid Cystic Carcinomas (ACCs) represent one of the most common and biologically aggressive forms of epithelial malignancy of the salivary glands, for which there are no clinically approved treatments. ACCs are characterized by a high tendency towards peri-neural infiltration and distant-site metastasis, and are largely refractory to conventional chemo-radiotherapy approaches. ACCs are also defined by a distinctive biological feature: the co-existence within malignant tissues of two phenotypically distinct sub-populations of cancer cells (ductal-like vs. myoepithelial-like) characterized by profoundly distinct gene-expression profiles and functional properties. This study will leverage a portfolio of surface markers recently identified by our laboratory that allow, for the first time, the differential purification, molecular study and functional analysis of these two cell types. The hypotheses formally tested by this study are: a) that the two cell types co-exist in a dynamic and hierarchical equilibrium, whereby one cell type functions as a progenitor for the other; and b) that biochemical pathways regulating the conversion of one cell type into the other can be leveraged to develop new therapeutic approaches against ACCs. The project envisions two specific aims: 1) to elucidate the developmental relationship between the two cellular components of ACCs: we will use fluorescence activated cell sorting (FACS) to purify the two sub-populations of cancer cells from human ACCs, and then employ prospective in vivo xenotransplantation experiments and single-cell RNA-seq technologies to test whether the two cell types: a) display robust and systematic differences in tumor-initiating capacity across patient-derived xenograft (PDX) lines from independent patients; b) are hierarchically related (i.e. whether both can plastically inter-covert into each other or whether one serves as the progenitor of the other, in a mono-directional fashion); c) contain additional, previously unrecognized cellular sub-populations, themselves characterized by distinct molecular and functional properties; 2) to test the in vitro and in vivo therapeutic efficacy of novel drug combinations against ACCs, designed to target biochemical pathways shown to affect the differential representation and preferential survival of the two cellular sub-types: we will treat 3D organoids and PDX lines shown to recapitulate the dual cell composition of primary ACCs with sequential drug regimens, designed to, in a first step, induce the differentiation of myoepithelial-like cells into ductal-like cells and, in a second step, to selectively eliminate ductal-like cells; such novel drug regimens will be tested for their capacity to: a) cause changes in the cell composition of malignant tissues (ductal/myoepithelial cell ratio); b) reduce tumor size and/or growth kinetics; c) prevent in vivo metastatic dissemination (using non-invasive bio-luminescent reporters). Significance. This study will elucidate the molecular and functional properties of the two cell types found in human ACCs, filling a major scientific gap in salivary tumor biology, and addressing an unmet clinical need in salivary gland oncology: the identification of novel drug targets for the pharmacological treatment of ACCs.