Faculty Bibliography

Angiogenesis is essential to wound repair, and vascular endothelial growth factor (VEGF) is a potent factor to stimulate angiogenesis. Here, we examine the potential of VEGF-overexpressing adipose-derived stromal cells (ASCs) for accelerating wound healing using nonviral, biodegradable polymeric vectors. Mouse ASCs were transfected with DNA plasmid encoding VEGF or green fluorescent protein (GFP) using biodegradable poly (beta-amino) esters (PBAE). Cells transfected using Lipofectamine 2000, a commercially available transfection reagent, were included as controls. ASCs transfected using PBAEs showed enhanced transfection efficiency and 12-15-fold higher VEGF production compared with cells transfected using Lipofectamine 2000 (*P < 0.05). When transplanted into a mouse wild-type excisional wound model, VEGF-overexpressing ASCs led to significantly accelerated wound healing, with full wound closure observed at 8 days compared to 10-12 days in groups treated with ASCs alone or saline control (*P < 0.05). Histology and polarized microscopy showed increased collagen deposition and more mature collagen fibers in the dermis of wound beds treated using PBAE/VEGF-modified ASCs than ASCs alone. Our results demonstrate the efficacy of using nonviral-engineered ASCs to accelerate wound healing, which may provide an alternative therapy for treating many diseases in which wound healing is impaired.

PGRN, a pleiotrophic growth factor, is known to play an important role in the maintenance and regulation of the homeostatic dynamics of normal tissue development, proliferation, regeneration, and the host-defense response and therefore, has been widely studied in the fields of infectious diseases, wound healing, tumorigenesis, and neuroproliferative and degenerative diseases. PGRN has also emerged as a multifaceted immune-regulatory molecule through regulating the signaling pathways known to be critical for immunology, especially TNF/TNFR signaling. In this review, we start with updates about the interplays of PGRN with ECM proteins, proteolytic enzymes, inflammatory cytokines, and cell-surface receptors, as well as various pathophysiological processes involved. We then review the data supporting an emerging role of PGRN in the fields of the "Cubic of I", namely, immunity, infection, and inflammation, with special focus on its regulation of autoimmune syndromes. We conclude with insights into the immunomodulating, anti-inflammatory, therapeutic potential of PGRN in treating diseases with an inflammatory etiology in a vast range of medical specialties.

Introduction: Portal/mesenteric vein resection (PVR) is technically challenging and adds potential morbidity to a pancreaticoduodenectomy (PD). We reviewed our experience with PD for pancreatic adenocarcinoma to evaluate both short and long term outcomes following PVR. Methods: From our institutional pancreatic adenocarcinoma database, we identified 223 patients who underwent pancreaticoduodenectomy (PD) with (Group I n= 20) or without (Group II n= 203) PVR during the period 1990-2011. The study end-points were overall morbidity, 30-day mortality, length of post-operative stay (LOS), overall survival (OS). Differences between groups were evaluated using t-test or chi-squared test. OS for each group was estimated with Kaplan-Meier method and compared using the log-rank statistics. Results: The two groups were similar in terms of gender, age, ethnicity, underlying comorbidities and performance status (see table 1). One patient in Group I and 8 in Group II were deemed borderline resectable (5.0% vs. 3.9%, p = 0.8) and underwent neo-adjuvant treatment. Duration of surgery was longer in Group I (532 vs. 456 min, p = 0.04), but there were no differences in operative blood losses (1047 vs. 991 ml, p = 0.8), length of stay (13.9 vs. 14.4 days, p = 0.8), overall morbidity (55% vs. 38%, p = 0.14). There were only 2 post-operative deaths, both in the Group II (p = 0.7). Pathology revealed similar TNM stage and rates of resections with negative margins (85% vs. 75%, p = 0.8). At median follow-up of 14 months there was no significant difference in OS (20.5 vs. 15.8 months, p = 0.6) Conclusions: In our experience, post-operative and long-term outcomes were not adversely affected by PVR. PVR should be offered to patients with pancreatic cancer involving portal or mesenteric veins. (Table Presented)

Mebendazole (MBZ) was identified as a promising therapeutic on the basis of its ability to induce apoptosis in melanoma cell lines through a B-cell lymphoma 2 (BCL2)-dependent mechanism. We now show that in a human xenograft melanoma model, oral MBZ is as effective as the current standard of care temozolomide in reducing tumor growth. Inhibition of melanoma growth in vivo is accompanied by phosphorylation of BCL2 and decreased levels of X-linked inhibitor of apoptosis (XIAP). Reduced expression of XIAP on treatment with MBZ is partially mediated by its proteasomal degradation. Furthermore, exposure of melanoma cells to MBZ promotes the interaction of SMAC/DIABLO with XIAP, thereby alleviating XIAP's inhibition on apoptosis. XIAP expression on exposure to MBZ is indicative of sensitivity to MBZ as MBZ-resistant cells do not show reduced levels of XIAP after treatment. Resistance to MBZ can be reversed partially by siRNA knockdown of cellular levels of XIAP. Our data indicate that MBZ is a promising antimelanoma agent on the basis of its effects on key antiapoptotic proteins.

Introduction: Increasing evidence suggests that the ratio of number of nodes harboring metastatic cancer to the total number of lymph node examined (lymph node ratio, LNR) affects survival after pancreatic resection for adenocarcinoma. We reviewed impact of lymph node status and LNR in our population of patients undergoing pancreatic resection for adenocarcinomMa.e thods: From our institutional pancreatic adenocarcinoma database, we identified 273 patients who underwent pancreatectomy during the period 1990-2011. of those, 51 had no nodes harvested in the specimen (No LN) and 86 had negative nodes (N0). Among those with positive nodes LNR wa<=s 0.1 in 27, <= 0.2 in 30, <= 0.3 in 21, <= 0.4 in 18 and > 0.4 in 40. Overall median survival was the study end point. Results: The 7 groups were similar in terms of gender, age, ECOG, primary procedure, and status of resection margins (see Table 1). T stage was higher in patients with elevated LNR (p=0.02). Survival was lower for patients with positive nodes (p < 0.01). This difference remained significant when excluding from analysis patients without harvested nodes (p = 0.005). Patient with LNR <= 0.1 had survival similar to N0 patients (20.1 vs. 20.0, p = 0.09). We observed a trend toward a worse survival in patients with higher LNR, which did not reach statistical significanCcoen. clusions: In our experience patients with LNR < 0.1 appeared to have survival similar to those with negative nodes. However LNR did not improve survival prognostication across patients with positive nodes. (Table Presented)

The transcription factor Nrf2 is responsible for regulating a battery of antioxidant and cellular protective genes, primarily in response to oxidative stress. A member of the cap 'n' collar family of transcription factors, Nrf2 activation is tightly controlled by a series of signaling events. These events can be separated into the basal state, a preinduction response, gene induction, and finally a postinduction response, culminating in the restoration of redox homeostasis. However, despite the immensely intricate level of control the cellular environment imposes on Nrf2 activity, there are many opportunities for perturbations to arise in the signaling events that favor carcinogenesis and, therefore, implicate Nrf2 as both a tumor suppressor and a protooncogene. Herein, we highlight the ways in which Nrf2 is regulated, and discuss some of the Nrf2-inducible antioxidant (NQO1, NQO2, HO-1, GCLC), antiapoptotic (Bcl-2), metabolic (G6PD, TKT, PPARgamma), and drug efflux transporter (ABCG2, MRP3, MRP4) genes. In addition, we focus on how Nrf2 functions as a tumor suppressor under normal conditions and how its ability to detoxify the cellular environment makes it an attractive target for other oncogenes either via stabilization or degradation of the transcription factor. Finally, we discuss some of the ways in which Nrf2 is being considered as a therapeutic target for cancer treatment.

Netrins have been extensively studied for their role in axonal guidance during neural development. In addition, netrins are chemopulsants for a variety of non-neuronal cell types via binding to their receptors Unc5b and DCC. Although thought to suppress inflammation in several settings, netrin1, acting via Unc5b, inhibits macrophage migration directed by chemokines CCL2 and CCL19 to promote macrophage retention in and exacerbation of atherosclerotic plaque. We asked whether Netrin1 was expressed during osteoclast (OC) differentiation and whether it plays a role in OC differentiation.DXAscan and MicroCT analysis were performed on Netrin1 deficient mice (radiation chimeras) and wildtype (WT, radiation chimeras) littermates. OC differentiation was studied as M-CSF/RANKL-stimulated differentiation of murine bone marrow precursors to TRAP+/multinucleated cells, in the presence/absence of recombinant Netrin1 and Unc5b antibody. Netrin1, Unc5b and DCC expression were studied by RT-PCR and Western Blot in primary bone marrow-derived osteoclasts. Netrin1 immunostaining was performed in human tissue obtained following primary prosthesis implantation or after prosthesis revision. During OC differentiation cell-associated Netrin1 and Unc5b (but not DCC) protein expression increased by 30+2% and 98+4% respectively (p<0.001,n=4) and Netrin1 secretion increased by 66+2% (p<0.001,n=4). Consistently, RANKL stimulates an increase in Netrin1 and Unc5b mRNA expression during OC differentiation (25+4 and 3+0.5 fold change respectively p<0.001,n=4). Moreover, in Netrin1-deficient marrow precursors OC differentiation was diminished by 65+2% as compared to control (p<0.001,n=6), an effect reversed by addition of recombinant netrin1 to cultures (121+5% increased, p<0.5,n=4). An antibody to the netrin1 receptor Unc5b reduces OC formation by 57+6% (p<0.001, =6) whereas an antibody to DCC had no effect on OC formation (5+4% reduction, p=NS vs. control,n=6). Finally, DXAscan and MicroCT analysis demonstrated an increase in!

Alterations in the metabolic control of lipid and glucose homeostasis predispose an individual to develop cardiometabolic diseases, such as type 2-diabetes mellitus and atherosclerosis. Work over the last years has suggested that microRNAs (miRNAs) play an important role in regulating these physiological processes. The contribution of miRNAs in regulating metabolism is exemplified by miR-33, an intronic miRNA encoded in the Srebp genes. miR-33 controls cellular cholesterol export and fatty acid degradation, whereas its host genes stimulate cholesterol and fatty acid synthesis. Other miRNAs, such as miR-122, also play a critical role in regulating lipid homeostasis by controlling cholesterol synthesis and lipoprotein secretion in the liver. This review article summarizes the recent findings in the field, highlighting the contribution of miRNAs in regulating lipid and glucose metabolism. We will also discuss how the modulation of specific miRNAs may be a promising strategy to treat metabolic diseases.

Progranulin (PGRN) is a multifunctional growth factor which is composed of seven-and-a-half repeats of a cysteine-rich motif in the order P-G-F-B-A-C-D-E. We previously reported that PGRN directly bound to TNF-alpha receptors (TNFR), and inhibited TNF-alpha activity (Tang, W, et al, Science. 2011). Atsttrin, an engineered protein composed of three PGRN fragments (1/2F-1/2A-1/2C), exhibited selective TNFR binding, and potently prevented inflammation in multiple arthritis mouse models (Tang, W, et al, Science. 2011). It is well established that TNF family ligands bind to receptors in a heterohexameric 3:3 complex. We created a novel engineering protein, Atsttrin primer, which comprised the same fragments as Atsttrin but in a different sequence (1/2A-1/2C-1/2F). The purpose of this study is to determine (1) whether the three fragments of Atsttrin (i.e. 1/2A, 1/2C and 1/2F) act independently for interacting with TNFR; and if so, (2) whether Atsttrin primer is able to block the binding of TNF-alpha to TNFR, and has therapeutic effect in inflammatory arthritis, as does Atsttrin. Molecular mechanistic studies through Yeast-two hybrid screening and Solid-phase binding assay revealed that Atsttrin primer had stronger binding affinity to TNFR compared with Atsttrin, and effectively inhibited TNF-alpha from binding to TNFRs. To define the role of Atsttrin primer in inflammatory arthritis, we induced collagen-induced-arthritis (CIA) model in