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.
These exploratory studies are relevant to public health for two reasons. First, this may provide a three dimensional microenvironment with flexibility to culture patient-specific lymphoma cells for which no robust platform exists. Second, they may provide accurate and efficient prediction on how epigenetic and metabolic reprogramming drugs should be administered to lymphoma patients and what sub-set of patients will more likely benefit from these agents.
|Singh, Ankur; Peppas, Nicholas A (2014) Hydrogels and scaffolds for immunomodulation. Adv Mater 26:6530-41|
|Patel, Ravi Ghanshyam; Purwada, Alberto; Cerchietti, Leandro et al. (2014) Microscale Bioadhesive Hydrogel Arrays for Cell Engineering Applications. Cell Mol Bioeng 7:394-408|