Project description:Purpose: To identify transcriptional changes by RNA-seq in tumor samples, before bevacizumab combination treatment and after bevacizumab combination treatment in both responding and non-responding recurrent glioblastoma patients
Project description:<p>The adoptive transfer of autologous T cells genetically modified to express a CD19-specific, 4-1BB/CD3zeta-signaling chimeric antigen receptor (CAR; CTL019) has shown remarkable activity in patients with B acute lymphoblastic leukemia. Similar therapy can induce long-term remissions for relapsed/refractory chronic lymphocytic leukemia (CLL) patients, but in only a small subset of subjects. The determinants of response and resistance to CTL019 therapy of CLL are not fully understood. We employed next generation sequencing of RNA (RNA-seq) to identify predictive indicators of response to CTL019 treatment. We performed RNA-seq on leukapheresis and manufactured infusion product T cells from patients with heavily pre-treated and high-risk disease. To characterize potency, we also performed RNA-seq on the cellular infusion product after CAR-specific stimulation. Our findings indicate that durable remission in CLL is associated with gene expression signatures of early memory T cell differentiation (e.g., STAT3), while T cells from poorly- or non-responding patients exhibited elevated expression of key regulators of late memory as well as effector T cell differentiation, apoptosis, aerobic glycolysis, hypoxia and exhaustion. These gene expression signatures, along with additional immunological biomarkers, may be used to identify which patients are most likely to respond to cellular therapies and suggest manufacturing modifications that might potentiate the generation of maximally efficacious infusion products.</p>
Project description:In mouse bone marrow, mesenchymal stem cells (MSC) has the potential to form osteocytes, adipocytes and cartilage. In the process of osteogenesis, MSCs differenetiate into stromal cells, such as CAR cells. Osteoblast is responsible for the formation of osteocytes and osteoblasts may be differentiated from a subset of CAR cells. Dmp1-Cre targeted CAR cells are thought to enrich for a osteoblast progenitor population. We used microarrays to detail the gene expression profiles among Dmp1-Cre targeted and non-targeted CAR cells. Gene expression diffferences were compared to support the hypothesis that Dmp1-Cre targeted CAR cells may be enriched for osteoblast progenitors. Dmp1-Cre targeted and non-targeted CAR cells were FACS sorted from three mice. RNA were extracted from these sorted cells and processed for microarray using Affymetrix mogene 1.0 ST chip. Cells from one mouse represent one sample