Efficacy assessment of interferon-alpha-engineered mesenchymal stromal cells in a mouse plasmacytoma model

Bone marrow mesenchymal stromal cells (BM-MSCs) may survive and proliferate in the presence of cycling neoplastic cells. Exogenously administered MSCs are actively incorporated in the tumor as stromal fibroblasts, thus competing with the local mesenchymal cell precursors. For this reason, MSCs have been suggested as a suitable carrier for gene therapy strategies, as they can be genetically engineered with genes encoding for biologically active molecules, which can inhibit tumor cell proliferation and enhance the anti-tumor immune response. We used BM-MSCs engineered with the murine interferon-alpha (IFN-alpha) gene (BM-MSCs/IFN-alpha) to assess in a mouse plasmacytoma model the efficacy of this approach towards neoplastic plasma cells. We found that IFN-alpha can be efficiently produced and delivered inside the tumor microenvironment. Subcutaneous multiple administration of BM-MSCs/IFN-alpha significantly hampered the tumor growth in vivo and prolonged the overall survival of mice. The anti-tumor effect was associated with enhanced apoptosis of tumor cells, reduction in microvessel density, and ischemic necrosis. By contrast, intravenous administration of BM-MSCs/IFN-alpha did not significantly modify the survival of mice, mainly as a consequence of an excessive entrapment of injected cells in the pulmonary vessels. In conclusion, BM-MSCs/IFN-alpha are effective in inhibiting neoplastic plasma cell growth; however, systemic administration of engineered MSCs still needs to be improved to make this approach potentially suitable for the treatment of multiple myeloma

Functional MRI changes in the central motor system in myotonic dystrophy type 1

Myotonic dystrophy type 1 (DM1) is a multisystemic disease involving multiple organ systems including central nervous system (CNS) and muscles. Few studies have focused on the central motor system in DM1, pointing to a subclinical abnormality in the CNS. The aim of our study was to investigate patterns of cerebral activation in DM1 during a motor task using functional MRI (fMRI). Fifteen DM1 patients, aged 20 to 59 years, and 15 controls of comparable age were scanned during a self-paced sequential finger-to-thumb opposition task of their dominant right hand. Functional MRI images were analyzed using SPM99. Patients underwent clinical and genetic assessment; all subjects underwent a conventional MR study. Myotonic dystrophy type 1 patients showed greater activation than controls in bilateral sensorimotor areas and inferior parietal lobules, basal ganglia and thalami, in the ipsilateral premotor area, insula and supplementary motor area (corrected P<05). Analysis of the interaction between disease and age showed that correlation with age was significantly greater in patients than in controls in bilateral sensorimotor areas and in contralateral parietal areas. Other clinical and MR characteristics did not correlate with fMRI. Functional changes in DM1 may represent compensatory mechanisms such as reorganization and redistribution of functional networks to compensate for ultrastructural and neurochemical changes occurring as part of the accelerated aging process

MR venography in patients with multiple sclerosis and correlation with clinical and MRI parameters

BACKGROUND AND PURPOSE: Multiple sclerosis (MS) has been associated with chronic cerebrospinal venous insufficiency. We aim to evaluate the correlation between extracranial veins stenosis evaluated with MR venography (MRV) and clinical/MR parameters of MS. METHODS: In 29 consecutive MS patients we performed a standard brain MRI protocol, completed by the evaluation of extra-cerebral venous system using a phase-contrast and a Volumetric Interpolated Breath Hold Examination (VIBE) sequence before and after gadolinium. The T2-proton density images were used to calculate the lesion volume. The jugular veins were evaluated qualitatively (in terms of presence and severity of stenoses) and quantitatively (degree of stenosis). The phase-contrast images were analyzed to calculate the average and peak velocity in the internal jugular veins. RESULTS: Postcontrast VIBE successfully showed the jugular veins in all the subjects. T2-lesion-volume was 8.2 [4.6] cm³. A stenosis of the internal jugular veins > of 50% was observed in 10/29(33%) patients. No significant correlation was observed between T2-lesion-volume and degree-of-stenosis (r = .362, P = .302). No different flow parameters were found in the subgroups of patients with and without stenosis (P = .54). CONCLUSIONS: In MS the presence/severity of jugular vein stenosis identified with 3T-MRV is not related to MR-visible tissue damage. Moreover no abnormal flow parameters were found in stenosed veins

Interferon-alpha-engineered Multipotent Mesenchymal Stromal Cells for the treatment of myeloma

Bone marrow mesenchymal stromal cells (BM-MSCs) are nonhematopoietic progenitor cells with multilineage differentiation potential. Exogenously administered BM-MSCs have been shown to survive and proliferate in the presence of malignant cells, becoming stromal cells and supporting tumor growth. Thus, BM-MSCs are attractive candidates to deliver biologically active molecules in the tumor environment in vivo and to enhance specific immune responses. Interferon-a (IFN-a) has been used for years for the maintenance treatment of multiple myeloma (MM), but its administration is limited by the temporary efficacy and the toxicity. We analyzed the in vivo effects of mouse BM-MSCs transduced with IFN-a cDNA in the Sp6 plasmacytoma mouse model. BM-MSCs were transduced with a lentiviral vector containing EGFP cDNA or murine IFN-a cDNA (efficiency = 70%). Two months-old Balb/c mice (Balb/cByJIco, Charles River Italia, Calco, LC, Italy) (H-2d), were injected subcutaneously (s.c.) with the tumorigenic dose of 0.5x106 Sp6 cells (H-2d). The same mice were then weekly injected with 0.5x106 BMMSCs/ IFN-a (1, 4 or 8 doses), in the same anatomical quarter. Some mice was injected s.c. with Sp6 and with BM-MSCs/EGFP s.c. or intravenously (i.v.) to test in vivo homing. Tumor immunohistochemistry was performed with anti-von Willebrand factor, anti- a -smooth muscle actin, anti-CD4, anti-CD8, anti-asialo GM1, anti-CD45, anti-CD90, anti-murine IFN-a. BM-MSCs were capable of homing into Sp6 tumor, forming clusters of cells. Treatment with BM-MSCs/IFN-a resulted in a significant delay in the onset of palpable tumors (event free survival, EFS, of 50% at day +17 for 1 dose, +20 for 4 doses and +64, for 8 doses, whereas Sp6 alone or coinjected with BM-MSCs showed tumor incidence of 100% 10-13 days after injection). Weekly delivery of BMMSCs/ IFN-a induced a significant decrease of kinetics tumor growth and an increment of overall survival (OS) (median OS in controls: 19 days, animals receiving BM-MSCs: 17 days, mice treated with 1 and 4 doses of BM-MSCs/IFN-a: 30-31 days, mice treated with 8 doses:77 days). The antitumor effect is associated with tumor necrosis, reduction in microvessel density, and NK cell infiltration. These findings indicate that transduced BM-MSCs could be useful to deliver anti-cancer molecules in the microenvironment of myeloma and become a promising tool for specific, low-toxic, and long-lasting anti-myeloma therapy

Bisdemethoxycurcumin (BDC)-Loaded H-Ferritin-Nanocages Mediate the Regulation of Inflammation in Alzheimer’s Disease Patients

Background: Bisdemethoxycurcumin (BDC) might be an inflammation inhibitor in Alzheimer’s Disease (AD). However, BDC is almost insoluble in water, poorly absorbed by the organism, and degrades rapidly. We thus developed a new nanoformulation of BDC based on H-Ferritin nanocages (BDC-HFn). Methods: We tested the BDC-HFn solubility, stability, and ability to cross a blood–brain barrier (BBB) model. We tested the effect of BDC-HFn on AD and control (CTR) PBMCs to evaluate the transcriptomic profile by RNA-seq. Results: We developed a nanoformulation with a diameter of 12 nm to improve the solubility and stability. The comparison of the transcriptomics analyses between AD patients before and after BDC-HFn treatment showed a major number of DEG (2517). The pathway analysis showed that chemokines and macrophages activation differed between AD patients and controls after BDC-HFn treatment. BDC-HFn binds endothelial cells from the cerebral cortex and crosses through a BBB in vitro model. Conclusions: Our data showed how BDC-Hfn could improve the stability of BDC. Significant differences in genes associated with inflammation between the same patients before and after BDC-Hfn treatment have been found. Inflammatory genes that are upregulated between AD and CTR after BDC-HFn treatment are converted and downregulated, suggesting a possible therapeutic approach

Plasma Lipid Profiling Contributes to Untangle the Complexity of Moyamoya Arteriopathy

Moyamoya arteriopathy (MA) is a rare cerebrovascular disorder characterized by ischemic/hemorrhagic strokes. The pathophysiology is unknown. A deregulation of vasculogenic/angiogenic/inflammatory pathways has been hypothesized as a possible pathophysiological mechanism. Since lipids are implicated in modulating neo-vascularization/angiogenesis and inflammation, their deregulation is potentially involved in MA. Our aim is to evaluate angiogenic/vasculogenic/inflammatory proteins and lipid profile in plasma of MA patients and control subjects (healthy donors HD or subjects with atherosclerotic cerebrovascular disease ACVD). Angiogenic and inflammatory protein levels were measured by ELISA and a complete lipidomic analysis was performed on plasma by mass spectrometry. ELISA showed a significant decrease for MMP-9 released in plasma of MA. The untargeted lipidomic analysis showed a cumulative depletion of lipid asset in plasma of MA as compared to HD. Specifically, a decrease in membrane complex glycosphingolipids peripherally circulating in MA plasma with respect to HD was observed, likely suggestive of cerebral cellular recruitment. The quantitative targeted approach demonstrated an increase in free sphingoid bases, likely associated with a deregulated angiogenesis. Our findings indicate that lipid signature could play a central role in MA and that a detailed biomarker profile may contribute to untangle the complex, and still obscure, pathogenesis of MA

Change in eGFR during FU according to TAp in all patients and in patients with eGFR <30 ml/min.

<p>Change in eGFR during FU according to TAp in all patients and in patients with eGFR <30 ml/min.</p

Renal survival.

<p>(A) Kaplan-Meier estimates of proportion of renal survival (absence of creatinine doubling) in the 325 patients by TAp groups. Estimated 5-year overall survival rates were 97.7% (95% CI 84.9–99.7) for patients with <0.3 g/day; 95.1% (95% CI 89.4–97.8) for patients with 0.3–0.9 g/day, 92.1% (95% CI 79.8–96.3) for those with 1–1.9 g/day, 69.4% (95% CI 46.3–84.1) for those with 2–2.9 g/day and 29.0% (95% CI 14.6–45.1) for those with ≥3 g/day. (B) Kaplan-Meier estimates of proportion of renal survival (absence of ESRD) in the 325 patients by TAp groups. Estimated 5-year overall survival rates were 100% (95% CI: not calculable) for patients with <0.3 g/day; 96.5% (95% CI 90.1–98.7) for patients with <0.3–0.9 g/day, 95.6% (95% CI 84.3–99.2) for those with 1–1.9 g/day, 69.1% (95% CI 45.6–84.0) for those with 2–2.9 g/day and 46.9% (95% CI 27.7–63.9–45.1) for those with ≥3 g/day.</p

End-stage (39 patients): risk factors at univariate analysis (Cox model with multiple imputation).

<p>End-stage (39 patients): risk factors at univariate analysis (Cox model with multiple imputation).</p