Bioengineered Lung Tumor Organoids For Development Of Personalized Medicine

 描述（由申请人提供）：肺癌影响美国大量人群，长期预后非常差。非小细胞肺癌（NSCLC）不再是一种单一的疾病，而是一系列根据组织学病理分类的癌症类型，这些癌症对药物的反应不同。因此，个性化/精确肿瘤学被提议作为NSCLC和其他癌症治疗的标准临床实践。这种对肿瘤DNA进行测序以鉴定可操作基因突变的实践依赖于足够量的完整肿瘤细胞DNA的可用性，并且产生了开发组织活检检索、处理和分析的高保真过程的需要。然而，没有统一的方法来满足这一需求，并且非常小和低纯度的肿瘤，例如肺的显微转移和细针抽吸（FNA）活检，在获得癌细胞特异性DNA方面存在固有的挑战，因此可能会使患者无法享受精准医疗的益处。或者，扩增活检衍生的细胞可以解决这个问题。这一提议的动机是迫切需要了解肿瘤活检的细胞扩增过程在多大程度上可能对下游分子和细胞分析产生负面影响-这些影响充其量也难以检测和消除。一般的癌症研究，特别是原代癌细胞的扩增，仍然依赖于使用塑料培养皿的标准细胞培养技术;因此，为细胞提供人工培养条件，对细胞施加选择性压力，这可能会大大改变其原始分子特性。我们假设，通过重现体内肺微环境，我们将能够成功地在体外扩增少量新鲜分离的肺癌细胞，同时保留其细胞和遗传表型，包括其突变谱。为了验证这一假设，我们提出生物打印生物工程肺类器官（BLO），由肺内皮细胞和成纤维细胞组成，包埋在肺特异性细胞外基质（ECM）内，并在肺肿瘤类器官（BLTO）内扩增肺肿瘤细胞。未来的发展可能包括患者特异性BLTO作为测试个性化治疗效率的替代物，BLTO也可用于阐明肿瘤生长和侵袭的新/新颖机制并鉴定新的治疗靶点。

 DESCRIPTION (provided by applicant):Lung cancer affects a large number of people in the U.S. with very poor long-term prognosis. Non-small cell carcinoma of the lung (NSCLC) is no longer a single disease, but a constellation of cancer types pathologically classified by histology which respond differently to drugs. As a consequence, personalized/precision oncology is proposed as a standard clinical practice for NSCLC and other cancers treatment. This practice, whereby tumor DNA is sequenced to identify actionable gene mutations, is dependent on the availability of sufficient amounts of intact tumor cell DNA, and creates a need to develop a high fidelity process of tissue biopsy retrieval, processing and analysis. However, there are no uniform methods to address this need and very small and low-purity tumors, such as microscopic metastases of the lung and fine-needle aspirate (FNA) biopsies, present an inherent challenge in obtaining cancer cell-specific DNA, and thus may preclude patients from the benefits of precision medicine. Alternatively, expansion of biopsy-derived cells could address this problem. This proposal is motivated by the critical need to understand to what extent the process of cell expansion from tumor biopsy may negatively influence downstream molecular and cellular analyses - influences that, at best, are difficult to detect and remove. Cancer research in general, and specifically expansion of primary cancer cells, still relies on standard cell culture techniques that use plastic dishes; thus, presenting the cells with artificia culture conditions that impose a selective pressure on the cells that could substantially alter their original molecular properties. We hypothesize that by recapitulating the in vivo lung microenvironment we will be able to successfully expand a small number of freshly isolated lung cancer cells in vitro, while preserving their cellular and genetic phenotype, including their mutational profile. To test this hypothesis we propose to bioprint bioengineered lung organoids (BLOs), consisting of lung endothelial and fibroblastic cells, embedded inside lung-specific extracellular matrix (ECM), and expand lung tumor cells inside lung tumor organoids (BLTOs). Future developments may include patient-specific BLTOs as surrogates for testing the efficiency of the personalized treatments and BLTOs may also be used to elucidate new/novel mechanisms of tumor growth and invasion and identify new therapeutic targets.