Live Tumor Culture Core and Tissue Specific Culture System for Human Cancers

项目摘要 背景：癌症药物开发的许多步骤涉及使用标准癌细胞系（SCCL） 这些药物在许多靶向治疗的开发中发挥了重要作用，包括赫赛汀、格列卫和 他莫昔芬等。然而，建立模拟人类疾病的新CCL仍然具有挑战性， 体外克隆选择和原始肿瘤表型的丧失。此外，创造新细胞的效率 线非常低。因此，约90%的卵巢癌和乳腺癌研究是在5个CCL中进行的， 2.这些SCCL和白血病细胞系并不能完全代表人类疾病的真实多样性。 创新：细胞培养技术的一个持续存在的缺点是使用全血清、药物和 饲养层，其部分负责原始肿瘤表型的不良复制。为了 为了解决这些问题，我们开发了一系列无血清组织特异性培养（TSC）培养基， 建立新细胞系的效率> 50%，并保留了重要的组织特异性原始肿瘤特征。 初步数据：我们最近发表了TSC技术在卵巢癌（OvCa）中的首次应用， 描述了25种新的OvCa细胞系，其保留了患者肿瘤的分子、组织学和结果特征。 目标：我们的长期目标是开发新的TSC媒体和方法，将我们的技术扩展到所有人 肿瘤类型和改进的方法来解决间质和正常细胞过度生长的问题。 具体目标：虽然我们对所有肿瘤类型的工作，在这个补助金，我们专注于实体瘤（乳腺癌，卵巢癌）和肿瘤的治疗。 液体肿瘤（白血病）来说明我们的系统可以适于培养肿瘤类型的全谱。 目的1）新的乳腺癌细胞系的表征：我们配制了一种新的正常乳腺特异性的， 乳腺癌特异性培养基，我们将使用它来建立匹配的正常细胞系和癌细胞系。 目的2）新的急性髓系白血病（AML）细胞系的表征：我们配制了一种新的无血清 为AML定制的培养基，我们将使用它来建立新的AML细胞系。 目的3）次优样品的培养：具有<10%肿瘤的组织样品由于基质的限制而难以培养。 过度增长。我们开发了一种抑制基质的方法，以使次优组织样品的培养成为可能。 癌症相关生物标本科学技术的验证：基因组、转录组和蛋白质组 将每种细胞系的细胞密度与原始患者肿瘤、现有细胞系和肿瘤数据集进行比较。 重大改进和新能力。我们的TSC系统可以在没有饲养层的情况下维持细胞系， 药物、提取物和抑制基质和正常细胞的过度生长。 转型潜力。SCCL的生物学相关性非常可疑。因此， 使用SCCL开发的药物在临床上经常失败。我们提供生物相关的能力 BrCa、OvCa和AML细胞系可以对癌症药物开发产生变革性影响。

PROJECT SUMMARY Background: Many steps of cancer drug development involves the use of standard cancer cell lines (SCCL) which have been instrumental in the development of many targeted therapies; including Herceptin, Gleevac and Tamoxifen among others. However, establishing new CCL that mimic human disease is still challenging due to in vitro clonal selection and loss of the original tumor phenotype. In addition, the efficiency of creating new cell lines is very low. As a result, ~90% of ovarian and breast cancer research has been carried out in 5 CCL each 1, 2. These SCCL and leukemia cell lines do not fully represent the real-life diversity of human disease. Innovation: One of the persistent shortcomings of cell culture technology is the use of full serum, drugs and feeder layers that are partly responsible for the poor replication of the original tumor phenotype. In order to address these problems we developed a series of serum-free tissue specific culture (TSC) media that increases the efficiency of establishing new cell lines to > 50% and retains important tissue specific original tumor features. Preliminary data: We recently published the first application of the TSC technology for ovarian cancer (OvCa), describing 25 new OvCa cell lines that retain molecular, histologic and outcome features of the patient tumors. Objectives: Our long-term goal is to develop novel TSC media and methods to expand our technology to all tumor types and improve the methods to solve the stromal and normal cell overgrowth problem. Specific Aims: While we work on all tumor types, in this grant we focus on solid tumors (Breast, Ovary) and a liquid tumor (Leukemia) to illustrate that our system can be adapted to culture the full spectrum of tumor types. Aim 1) Characterization of new breast cancer cell lines: We formulated a new normal breast specific and breast cancer specific media, which we will use to establish matching normal and cancer cell lines. Aim 2) Characterization of new acute myeloid leukemia (AML) lines: We formulated a new serum-free medium customized for AML, which we will use to establish new AML cell lines. Aim 3) Culture of sub-optimal samples: Tissue samples with <10% tumor are difficult to culture due to stromal over-growth. We developed a method that suppress stroma to enable culture of sub-optimal tissue samples. Validation of Cancer-Relevant Biospecimen Science Technology: The genome, transcriptome and proteome of each cell line will be compared to the original patient tumor, existing cell lines and tumor datasets. Substantial Improvement and New Capabilities. Our TSC system can maintain cell lines without feeder layers, drugs, extracts, and suppresses the over-growth of stromal and normal cells. Transformative Potential. The biological relevance of SCCL is highly questionable. Hence, it is not surprising that drugs that are developed using SCCL frequently fail in the clinic. Our ability to provide biologically relevant BrCa, OvCa, and AML cell lines can have a transformative effect on cancer drug development.