Bioengineered Lung Tumor Organoids For Development Of Personalized Medicine

 描述(由申请者提供)：肺癌在美国影响着大量的人，长期预后非常差。非小细胞肺癌(NSCLC)不再是一种单一的疾病，而是根据组织学病理分类的对药物反应不同的一系列癌症类型。因此，个性化/精确肿瘤学被提议作为非小细胞肺癌和其他癌症治疗的标准临床实践。这种做法，即对肿瘤DNA进行测序以识别可操作的基因突变，依赖于足够数量的完整肿瘤细胞DNA的可用性，并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，非常小和低纯度的肿瘤，如肺的微观转移和细针抽吸(FNA)活检，在获得癌细胞特异性DNA方面提出了固有的挑战，因此可能使患者无法享受精确医学的好处。或者，活组织检查衍生细胞的扩增可以解决这个问题。这一建议的动机是迫切需要了解肿瘤活检的细胞扩张过程可能在多大程度上对下游分子和细胞分析产生负面影响--这些影响充其量是难以检测和消除的。总的来说，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，可能会显著改变其原始分子特性。我们假设，通过重述体内肺微环境，我们将能够成功地在体外扩增少量新分离的肺癌细胞，同时保留它们的细胞和遗传表型，包括它们的突变情况。为了验证这一假设，我们建议生物打印由肺内皮细胞和成纤维细胞组成的生物工程肺器官(BLOS)，将其嵌入肺特异性细胞外基质(ECM)中，并在肺肿瘤器官(BLTO)内扩增肺肿瘤细胞。未来的发展可能包括针对患者的BLTO作为测试个性化治疗效率的替代品，BLTO还可能用于阐明肿瘤生长和侵袭的新/新机制，并确定新的治疗靶点。

 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.