Armstrong, Adam

Abstract Introduction: Immune suppression is a major component of lymphoma pathogenesis. Recent advances in lymphoma treatment have been immunotherapy. We present the preliminary results of our clinical trial testing 2 vaccine strategies in patients with indolent B-cell non-Hodgkin lymphoma (NHL). The primary objective is to determine the safety and feasibility of the vaccine approaches and secondary objectives are to describe clinical responses and identify corresponding immune changes. Methods: Autologous mature dendritic cells (mDC) were manufactured from leukapheresed cells of NHL pts. For patients with tumor lymph nodes deemed amenable to cryoablation by interventional radiologist (arm A), they received cryoablation of a node and injection of mDC into the cryoablated node followed by another 1 to 7 intratumoral mDC injections. Remaining patients had a tumor excised to generate tumor lysate ex vivo. mDC were pulsed with tumor lysate during DC maturation (arm B; DC-TL). The DC-TL vaccines were injected intradermally for 4 to 8 doses. Patients are monitored for one year after vaccines for adverse events and systemic tumor response. Correlative studies include cellular immune phenotype of peripheral blood and T cell intracellular cytokine productions. Planned accrual is 10 patients per arm (total = 20). Results: To date, 10 patients have accrued to arm A and 5 patients to arm B (Table 1). All pts tolerated vaccine treatments without major adverse events. Of the 10 evaluable patients, there were 1 CR (arm B: 1 / 4; total: 1/6) and 4 PR (arm A: 3/6; arm B: 1 / 4; total: 4/10) for an ORR of 50% for both arms (Figure 1). Time to next treatment ranged from 6.5 to 25 months (Figure 2). Correlative studies suggest that immune changes can be used as prognostic biomarkers to predict response. These include changes in vaccine induced cytokine secretion, and in the number and composition of the immune system. In addition, preliminary analysis of >40 immune phenotypes using flow cytometry and hierarchical clustering suggest there are characteristic immune system responses in patients responding to vaccine. Conclusions: Both cryoablation and intratumoral mDC vaccination are feasible and safe in NHL. Treatment responses may correlate with immune system changes. Biosystems analysis method can be used to develop novel assays as predictive biomarkers of treatment response and used to develop novel protocols that allow treating to response. Table 1. Patient Demographics Arm A (n = 10) Arm B (n = 5) Total (n = 15) Age (median, rage) 64 (55 – 80) 60 (31 – 86) 63 (31 – 86) Gender (M/F) 6/4 2/3 8/7 Histology (n, %) Follicular Marginal zone DLBCL 10 (100%) 0 (0%) 0 (0%) 3 (60%) 1 (20%) 1(20%) 13 (86.6%) 1 (6.7%) 1 (6.7%) Stage (N, %) II III IV 1 (10%) 1 (10%) 8 (80%) 2 (40%) 1 (20%) 2 (40%) 3 (20%) 2 (13.3%) 10 (66.7%) No of prior Tx (mean, range) 2.4 (0-8) 3.2 (0-7) 2.7 (0-8) FLIPI (n, %) 0-1 2 ≥ 3 1 (10%) 2 (20%) 7 (70%) 1 (20%) 2 (40%) 2 (45%) 2 (13.3%) 4 (26.7%) 9 (60%) Positive GELF criteria (n, %) 4 (40%) 1 (20%) 5 (30%) Figure 1. Waterfall plot. Figure 1. Waterfall plot. Figure 2. Time to next treatment. Figure 2. Time to next treatment. Disclosures No relevant conflicts of interest to declare.

Atherosclerotic renovascular disease (RVD) reduces renal blood flow (RBF) and GFR and accelerates poststenotic kidney (STK) tissue injury. Preclinical studies indicate that mesenchymal stem cells (MSCs) can stimulate angiogenesis and modify immune function in experimental RVD. We assessed the safety and efficacy of adding intra-arterial autologous adipose-derived MSCs into STK to standardized medical treatment in human subjects without revascularization. The intervention group ( n =14) received a single infusion of MSC (1.0 × 10 5 or 2.5 × 10 5 cells/kg; n =7 each) plus standardized medical treatment; the medical treatment only group ( n =14) included subjects matched for age, kidney function, and stenosis severity. We measured cortical and medullary volumes, perfusion, and RBF using multidetector computed tomography. We assessed tissue oxygenation by blood oxygen level–dependent MRI and GFR by iothalamate clearance. MSC infusions were well tolerated. Three months after infusion, cortical perfusion and RBF rose in the STK (151.8–185.5 ml/min, P =0.01); contralateral kidney RBF increased (212.7–271.8 ml/min, P =0.01); and STK renal hypoxia (percentage of the whole kidney with R2*>30/s) decreased (12.1% [interquartile range, 3.3%–17.8%] to 6.8% [interquartile range, 1.8%–12.9%], P =0.04). No changes in RBF occurred in medical treatment only subjects. Single-kidney GFR remained stable after MSC but fell in the medical treatment only group (−3% versus −24%, P =0.04). This first-in-man dose-escalation study provides evidence of safety of intra-arterial infusion of autologous MSCs in patients with RVD. MSC infusion without main renal artery revascularization associated with increased renal tissue oxygenation and cortical blood flow.

Abstract The induced pluripotent stem cell (iPSC) technology enables derivation of patient-specific pluripotent stem cells from adult somatic cells without using an embryonic cell source. Redifferentiation of iPSCs from diabetic patients into pancreatic islets will allow patient-specific disease modeling and autologous cell replacement therapy for failing islets. To date, diabetes-specific iPSCs have been generated from patients with type 1 diabetes using integrating retroviral vectors. However, vector integration into the host genome could compromise the biosafety and differentiation propensities of derived iPSCs. Although various integration-free reprogramming systems have been described, their utility to reprogram somatic cells from patients remains largely undetermined. Here, we used nonintegrating Sendai viral vectors to reprogram cells from patients with type 1 and type 2 diabetes (T2D). Sendai vector infection led to reproducible generation of genomic modification-free iPSCs (SV-iPSCs) from patients with diabetes, including an 85-year-old individual with T2D. SV-iPSCs lost the Sendai viral genome and antigens within 8–12 passages while maintaining pluripotency. Genome-wide transcriptome analysis of SV-iPSCs revealed induction of endogenous pluripotency genes and downregulation of genes involved in the oxidative stress response and the INK4/ARF pathways, including p16INK4a, p15INK4b, and p21CIP1. SV-iPSCs and iPSCs made with integrating lentiviral vectors demonstrated remarkable similarities in global gene expression profiles. Thus, the Sendai vector system facilitates reliable reprogramming of patient cells into transgene-free iPSCs, providing a pluripotent platform for personalized diagnostic and therapeutic approaches for diabetes and diabetes-associated complications.

Streptozotocin (STZ)-induced diabetes mellitus (DM) offers a very cost-effective and expeditious technique that can be used in most strains of rodents, opening the field of DM research to an array of genotypic and phenotypic options that would otherwise be inaccessible. Despite widespread use of STZ in small animal models, the data available concerning drug preparation, dosing and administration, time to onset and severity of DM, and any resulting moribundity and mortality are often limited and inconsistent. Because of this, investigators inexperienced with STZ-induced diabetes may find it difficult to precisely design new studies with this potentially toxic chemical and account for the severity of DM it is capable of inducing. Until a better option becomes available, attempts need to be made to address shortcomings with current STZ-induced DM models. In this paper we review the literature and provide data from our pancreatic islet transplantation experiments using single high-dose STZ-induced DM in NCr athymic nude mice with hopes of providing clarification for study design, suggesting refinements to the process, and developing a more humane process of chemical diabetes induction.