Regression of established hepatocellular carcinoma is induced by chemoimmunotherapy in an orthotopic murine model

The high rate of mortality and frequent incidence of recurrence associated with hepatocellular carcinoma (HCC) reveal the need for new therapeutic approaches. In this study we evaluated the efficacy of a novel chemoimmunotherapeutic strategy to control HCC and investigated the underlying mechanism that increased the antitumor immune response. We developed a novel orthotopic mouse model of HCC through seeding of tumorigenic hepatocytes from SV40 T antigen (Tag) transgenic MTD2 mice into the livers of syngeneic C57BL/6 mice. These MTD2-derived hepatocytes form Tag-expressing HCC tumors specifically within the liver. This approach provides a platform to test therapeutic strategies and antigen-specific immune-directed therapy in an immunocompetent murine model. Using this model we tested the efficacy of a combination of oral sunitinib, a small molecule multitargeted receptor tyrosine kinase (RTK) inhibitor, and adoptive transfer of tumor antigen-specific CD8(+) T cells to eliminate HCC. Sunitinib treatment alone promoted a transient reduction in tumor size. Sunitinib treatment combined with adoptive transfer of tumor antigen-specific CD8(+) T cells led to elimination of established tumors without recurrence. In vitro studies revealed that HCC growth was inhibited through suppression of STAT3 signaling. In addition, sunitinib treatment of tumor-bearing mice was associated with suppression of STAT3 and a block in T-cell tolerance. CONCLUSION: These findings indicate that sunitinib inhibits HCC tumor growth directly through the STAT3 pathway and prevents tumor antigen-specific CD8(+) T-cell tolerance, thus defining a synergistic chemoimmunotherapeutic approach for HCC.     

Inhibition of hepatic sulfatase-2 in vivo: a novel strategy to correct diabetic dyslipidemia

Type 2 diabetes mellitus (T2DM) impairs hepatic clearance of atherogenic postprandial triglyceride-rich lipoproteins (TRLs). We recently reported that livers from T2DM db/db mice markedly overexpress the heparan sulfate glucosamine-6-O-endosulfatase-2 (SULF2), an enzyme that removes 6-O sulfate groups from heparan sulfate proteoglycans (HSPGs) and suppresses uptake of TRLs by cultured hepatocytes. In the present study, we evaluated whether Sulf2 inhibition in T2DM mice in vivo could correct their postprandial dyslipidemia. Selective second-generation antisense oligonucleotides (ASOs) targeting Sulf2 were identified. Db/db mice were treated for 5 weeks with Sulf2 ASO (20 or 50 mg/kg per week), nontarget (NT) ASO, or phosphate-buffered saline (PBS). Administration of Sulf2 ASO to db/db mice suppressed hepatic Sulf2 messenger RNA expression by 70%-80% (i.e., down to levels in nondiabetic db/m mice) and increased the ratio of tri- to disulfated disaccharides in hepatic HSPGs (P < 0.05). Hepatocytes isolated from db/db mice on NT ASO exhibited a significant impairment in very-low-density lipoprotein (VLDL) binding that was entirely corrected in db/db mice on Sulf2 ASO. Sulf2 ASO lowered the random, nonfasting plasma triglyceride (TG) levels by 50%, achieving nondiabetic values. Most important, Sulf2 ASO treatment flattened the plasma TG excursions in db/db mice after corn-oil gavage (iAUC, 1,500 ± 470 mg/dL·h for NT ASO versus 160 ± 40 mg/dL · h for Sulf2 ASO\P < 0.01). CONCLUSIONS: Despite extensive metabolic derangements in T2DM mice, inhibition of a single dys-regulated molecule, SULF2, normalizes the VLDL-binding capacity of their hepatocytes and abolishes postprandial hypertriglyceridemia. These findings provide a key proof of concept in vivo to support Sulf2 inhibition as an attractive strategy to improve metabolic dyslipidemia.     

Twenty-four-hour profiles of plasma glucose, insulin, C-peptide and free fatty acid in subjects with varying degrees of glucose tolerance following short-term, medium-dose prednisone (20 mg/day) treatment: evidence for differing effects on insulin secretion and action

Objective To determine the time course and prandial effects of short‐term, medium‐dose prednisone on 24‐h metabolic patterns under standardized conditions. Context Glucocorticoids (GCs) adversely affect glucose homoeostasis but 24‐h profiles of glucose, insulin, C‐peptide and free fatty acids (FFAs) following short‐term, medium‐dose prednisone treatment in persons with varying degrees of glucose tolerance are not well defined. Design An open‐label cross‐sectional interventional study. Subjects Three groups were prospectively studied: persons with type 2 diabetes (T2DM; n = 7), persons ‘at risk’ for T2DM (AR; n = 8) and persons with normal glucose tolerance (NGT; n = 5). Methods Before and after 3‐day treatment with prednisone 20 mg each morning, subjects underwent 24‐h frequent blood sampling. Eucaloric mixed meals were provided at 08:00, 12:00 and 18:00 h. Insulin/glucose ratio provided an estimate of β‐cell response to meal stimuli. Measurements Plasma glucose, insulin, C‐peptide, haemoglobin A1c and FFA. Results Prednisone induced greater increases in glucose levels from midday (P = 0·001) to midnight (P = 0·02) in the T2DM than the AR and NGT groups. In contrast, insulin (P = 0·03) and C‐peptide (P = 0·04) levels decreased postbreakfast in the T2DM group, whereas no changes in the morning but higher C‐peptide levels (P = 0·03) from midday to midnight were observed in the AR group. In the T2DM group, insulin/glucose ratio decreased postbreakfast (P = 0·04) and increased postdinner (P = 0·03). Fasting glucose, insulin and C‐peptide levels were unchanged in all groups, and FFA levels modestly increased postdinner (P = 0·03) in the NGT group. Conclusion Short‐term, medium‐dose prednisone treatment induces postprandial hyperglycaemia in T2DM and AR predominantly from midday to midnight because of suppression of insulin secretion followed by decreased insulin action that dissipates overnight. Effective treatment of prednisone‐induced hyperglycaemia should target both rapid onset relative insulin deficiency and a less than 24‐h total duration of effect.     

Coronary arteriovenous fistulae: the complexity of coronary artery-to-coronary sinus connections

Coronary artery fistulae are a rare cardiovascular anomaly. Even less common are multiple fistulae involving more than 1 coronary artery. Herein we report the case of a 47-year-old woman who had fistulae from both the right coronary and left circumflex coronary arteries draining into the coronary sinus. These lesions we attempted to close percutaneously but subsequently closed surgically. We discuss diagnostic imaging approaches, along with closure indications, closure options, and outcomes.