(PQD5)Biomaterials-based Adaptive Tumor Microenvironments for In Vitro Drug Scree

描述（由申请人提供）：非霍奇金淋巴瘤（NHL）是一种起源于B细胞和T细胞的淋巴细胞增殖性疾病，用化疗药物治疗，成功率不同，几十年来几乎没有改变。显然需要更具体、毒性更小的治疗方法。一种有希望的方法是使用表观遗传药物，通过将细胞重编程为更化学敏感的表型，靶向转录复合物和DNA甲基化。缺乏将这些药物转化为患者的部分原因是缺乏足够的模型来准确有效地识别候选药物并建立治疗计划。例如，没有滤泡性淋巴瘤或大多数非皮肤t - nhl的细胞系模型。因此，这些疾病的治疗经验上是从弥漫性大B细胞淋巴瘤转化而来的，其结果要差得多，而且与患者的肿瘤相关。鉴于微环境对NHL生物学和药物反应的重要性日益增加，有必要开发模拟NHL微环境的3D组织，并允许患者的细胞特异性设计灵活性。该项目的目标是开发一种人工3D细胞外基质（ECM），它将支持患者来源的原代B细胞和T-NHL细胞的生存和生长，将具有设计灵活性，并将适用于抗肿瘤重编程药物的筛选。在正在进行的实验中，我们发现淋巴瘤细胞通过整合素分子与ECM之间的串扰对其存活和耐药很重要。初步数据表明，¿v¿3是8种人T-NHL细胞的促生存因子和化疗耐药因子，代表了未成熟T-NHL和外周血T-NHL的谱。此外，在一项正在进行的I期临床试验中，利用30个DLBCL系、9个T-NHL系和从患者身上获得的原代NHL细胞，研究人员证实，表观遗传药物如RGD配体结合DNA甲基转移酶整合素抑制剂（DNMT）可以特异性杀死原代人DLBCL细胞。鉴于这些发现，本研究的工作假设是，三维培养平台，如所提出的具有整合素特异性的粘附、可生物降解水凝胶，将在原发性NHL和支持基质细胞中发出促生存因子的信号，促进基质网络重塑，并增强药物的扩散。这些假设将在以下具体目标的实验中得到解决：(1)工程水凝胶提供整合素特异性配体，以改善NHL原代细胞的生长；(2)评估替代肿瘤水凝胶用于检测NHL中染色质和代谢重编程药物的功效。如果成功，这项探索性研究将改变淋巴瘤细胞的培养方式，但更重要的是，将允许更快，更合理的筛选和翻译治疗方案。功能化的3D微尺度水凝胶将模拟肿瘤样异质微环境，以支持原发性人类淋巴瘤的长期生长，结合患者淋巴瘤的分子特征，将允许更好地预测这组具有挑战性且可能非常有效的药物的药物反应。

DESCRIPTION (provided by applicant): Non-Hodgkin lymphomas (NHL) are a heterogeneous group of lymphoproliferative disorders of B and T cell origin that are treated with chemotherapy drugs with variable success rate that has virtually not changed over decades. There is a clear need for more specific and less toxic treatments. One promising approach is the use of epigenetic drugs that target transcriptional complexes and DNA methylation, by reprogramming cells to a more chemosensitive phenotype. The paucity in translating these drugs to patients is, in part, due to the lack of adequate models to accurately and efficiently identify candidates and to build therapeutic schedules. For instance, there are no cell line models of follicular lymphoma or most non-cutaneous T-NHLs. Therefore, the treatment for these diseases is empirically translated from diffuse large B cell lymphomas, with much poorer outcomes and correlations to the patient's tumor. There is a need to develop 3D tissues that mimic the NHL microenvironment Given the increasing importance of the microenvironment to NHL biology and drug response, t and allow for a patient's cell specific design flexibility. The objective of this project is to devlop an artificial 3D extracellular matrix (ECM) that will support the survival and growth of patient-derived primary B and T-NHL cells, will possess design flexibility, and will be suitable for screening of anti-neoplastic reprogramming drugs. In ongoing experiments, it was found that the crosstalk between lymphoma cells and the ECM via integrin molecules is important for their survival and chemo-resistance. The preliminary data represents that ¿v¿3 is a pro-survival and chemoresistance factor for 8 human T-NHL cells representing the spectrum of immature and peripheral T-NHLs. Moreover, using 30 DLBCL lines, 9 T-NHL lines, and primary NHL cells obtained from patients in an ongoing Phase I clinical trial, it was established that epigenetic drugs like the RGD ligand binding integrin inhibitors of DNA Methyltransferase (DNMT) can specifically kill primary human DLBCL cells. Given these findings, the working hypothesis of the proposed study is that 3D culture platform such as the proposed adhesive, biodegradable hydrogels with integrin specificities will signal the pro-survival factors in primary NHL and supporting stromal cells, facilitate matrix network remodeling, and enhance diffusion of drugs. These hypotheses will be addressed in the experiments of the following Specific Aims: (1) Engineer hydrogels presenting integrin-specific ligands for improved growth of NH L primary cells, and (2) Evaluate the efficacy of surrogate tumor hydrogels for testing chromatin- and metabolic-reprogramming agents in NHL If successful, this exploratory study will change the way lymphoma cells are cultured, but more importantly, will allow a faster and more rational screening and translation of therapeutic regimens. The functionalized 3D microscale hydrogel will mimic a neoplasm-like heterogeneous microenvironment to support long-term growth of primary human lymphomas that, integrated to the molecular characterization of patients' lymphomas, will allow better drug response prediction of this challenging and potentially very effective group of drugs.