Cannabinoid receptor type 1 immunoreactivity and disease severity in human epithelial ovarian tumors.

Objective In light of recent findings indicating that endocannabinoid system has antitumor actions, our study aimed to localize it in the human epithelial ovarian tumors, highlighting the differences among benign, borderline, and invasive forms and correlating cannabinoid receptor type 1 (CB1R) expression with disease severity. Study Design We determined CB1R immunohistochemical expression in 66 epithelial ovarian tumors treated in the Department of Woman, Child, and General and Specialized Surgery, Second University of Naples, at S. Maria del Popolo degli Incurabili Hospital (Naples): 36 borderline ovarian tumors, the main target of interest being intermediate forms, 15 benign and 15 invasive ovarian tumors. Results The benign ovarian tumors showed a weak expression of CB1R in the 33% of the cases and moderate expression in the 67% of the cases. Borderline ovarian tumors had a similar trend. They showed weak CB1R expression in 28% of the cases, moderate expression in 53% of the cases, and strong expression in 19% of the cases. In contrast, invasive tumors showed a weak expression of CB1R in 7% of the cases, moderate expression in 20% of the cases, and strong expression in 73% of the cases. Conclusion The recorded data show that the expression of CB1R increased from benign and borderline to malignant tumors. In the near future, endocannabinoid receptors might be used in clinical practice, alone or in combination with other markers, to identify or better characterize ovarian tumors, without considering the great opportunity that they might represent as therapeutic targets

The early reduction of left ventricular mass after sleeve gastrectomy depends on the fall of branched-chain amino acid circulating levels

INTRODUCTION: Body-mass index is a major determinant of left-ventricular-mass (LVM). Bariatric-metabolic surgery (BMS) reduces cardiovascular mortality. Its mechanism of action, however, often encompasses a weight-dependent effect. In this translational study, we aimed at investigating the mechanisms by which BMS leads to LVM reduction and functional improvement. METHODS: Twenty patients (45.2 ± 8.5years) were studied with echocardiography at baseline and at 1,6,12 and 48 months after sleeve-gastrectomy (SG). Ten Wistar rats aged 10-weeks received high-fat diet ad libitum for 10 weeks before and 4 weeks after SG or sham-operation. An oral-glucose-tolerance-test was performed to measure whole-body insulin-sensitivity. Plasma metabolomics was analysed in both human and rodent samples. RNA quantitative Real-Time PCR and western blots were performed in rodent heart biopsies. The best-fitted partial-least-square discriminant-analysis model was used to explore the variable importance in the projection score of all metabolites. FINDINGS: Echocardiographic LVM (-12%,-23%,-28% and -43% at 1,6,12 and 48 months, respectively) and epicardial fat decreased overtime after SG in humans while insulin-sensitivity improved. In rats, SG significantly reduced LVM and epicardial fat, enhanced ejection-fraction and improved insulin-sensitivity compared to sham-operation. Metabolomics showed a progressive decline of plasma branched-chain amino-acids (BCAA), alanine, lactate, 3-OH-butyrate, acetoacetate, creatine and creatinine levels in both humans and rodents. Hearts of SG rats had a more efficient BCAA, glucose and fatty-acid metabolism and insulin signaling than sham-operation. BCAAs in cardiomyocyte culture-medium stimulated lipogenic gene transcription and reduced mRNA levels of key mitochondrial β-oxidation enzymes promoting lipid droplet accumulation and glycolysis. INTERPRETATION: After SG a prompt and sustained decrease of the LVM, epicardial fat and insulin resistance was found. Animal and in vitro studies showed that SG improves cardiac BCAA metabolism with consequent amelioration of fat oxidation and insulin signaling translating into decreased intra-myocytic fat accumulation and reduced lipotoxicity

Adenoviral gene transfer of a single-chain IL-23 induces psoriatic arthritis-like symptoms in NOD mice

Previously, we demonstrated that intratumoral delivery of adenoviral vector encoding single-chain (sc)IL-23 (Ad.scIL-23) was able to induce systemic antitumor immunity. Here, we examined the role of IL-23 in diabetes in nonobese diabetic mice. Intravenous delivery of Ad.scIL-23 did not accelerate the onset of hyperglycemia but instead resulted in the development of psoriatic arthritis. Ad.scIL-23-treated mice developed erythema, scales, and thickening of the skin, as well as intervertebral disc degeneration and extensive synovial hypertrophy and loss of articular cartilage in the knees. Immunological analysis revealed activation of conventional T helper type 17 cells and IL-17-producing gamma delta T cells along with a significant depletion and suppression of T cells in the pancreatic lymph nodes. Furthermore, treatment with anti-IL-17 antibody reduced joint and skin psoriatic arthritis pathologies. Thus, these Ad.scIL-23-treated mice represent a physiologically relevant model of psoriatic arthritis for understanding disease progression and for testing therapeutic approaches.-Flores, R. R., Carbo, L., Kim, E., Van Meter, M., De Padilla, C. M. L., Zhao, J., Colangelo, D., Yousefzadeh, M. J., Angelini, L. A., Zhang, L., Pola, E., Vo, N., Evans, C. H., Gambotto, A., Niedernhofer, L. J., Robbins, P. D. Adenoviral gene transfer of a single-chain IL-23 induces psoriatic arthritis-like symptoms in NOD mice

Adenoviral gene transfer of a single-chain IL-23 induces psoriatic arthritis-like symptoms in NOD mice.

Previously, we demonstrated that intratumoral delivery of adenoviral vector encoding single-chain (sc)IL-23 (Ad.scIL-23) was able to induce systemic antitumor immunity. Here, we examined the role of IL-23 in diabetes in nonobese diabetic mice. Intravenous delivery of Ad.scIL-23 did not accelerate the onset of hyperglycemia but instead resulted in the development of psoriatic arthritis. Ad.scIL-23–treated mice developed erythema, scales, and thickening of the skin, as well as intervertebral disc degeneration and extensive synovial hypertrophy and loss of articular cartilage in the knees. Immunological analysis revealed activation of conventional T helper type 17 cells and IL-17–producing γδ T cells along with a significant depletion and suppression of T cells in the pancreatic lymph nodes. Furthermore, treatment with anti–IL-17 antibody reduced joint and skin psoriatic arthritis pathologies. Thus, these Ad.scIL-23–treated mice represent a physiologically relevant model of psoriatic arthritis for understanding disease progression and for testing therapeutic approaches.—Flores, R. R., Carbo, L., Kim, E., Van Meter, M., De Padilla, C. M. L., Zhao, J., Colangelo, D., Yousefzadeh, M. J., Angelini, L. A., Zhang, L., Pola, E., Vo, N., Evans, C. H., Gambotto, A., Niedernhofer, L. J., Robbins, P. D. Adenoviral gene transfer of a single-chain IL-23 induces psoriatic arthritis–like symptoms in NOD mice. Psoriasis is a chronic inflammatory autoimmune disease of the skin that exists in 3% of the world population. Psoriasis vulgaris, or plaque psoriasis, which manifests as raised itchy erythematous plaques in the skin, is the most common form of psoriasis and accounts for nearly 70% of the patients with psoriasis (1). Psoriatic arthritis (PsA) is the second most common form of psoriasis and is found in ∼30% of patients with psoriasis (1). Patients with PsA have inflammation and autoimmunity occurring in the spine, the peripheral joints, and the site of attachment of ligament to the bone, as well as skin plaques (2). Patients with psoriasis are also prone to develop metabolic disorders, such as type 2 diabetes, cardiovascular disease, and kidney disease. Symptoms of psoriasis in humans include patches of red, inflamed skin with involved areas of the skin developing lesions, referred to as Koebner’s phenomenon. During the early development of psoriatic skin lesions, dermal changes include the formation of twisted dilated vessels along with papillar edema and an infiltrate mainly consisting of dendritic cells and macrophages (3, 4). Within the fully developed lesion, there is extensive thickening of the epidermis (acanthosis) with elongated epidermal rete ridges and dermal papillae with dilated capillaries. The epidermal layer also undergoes extensive hyperplasia, which leads to thickening of stratum corneum (hyperkeratosis), whereas the stratum granulosum decreases. This increase in the number of cells on the surface of the skin leads to thick, silvery scales, another hallmark of the disease. PsA is a chronic arthritic disease, ranging in severity from occasional flare-ups to continuous inflammation that can cause joint damage if not treated. PsA typically affects the large joints, especially those of the lower extremities and distal joints of the fingers. PsA also can affect the spine, leading to degeneration of intervertebral discs and the sacroiliac joints of the pelvis. Epidemiologic data show that in 60% of the cases, psoriatic skin lesions precede joint disease (2). IL-23, a member of the IL-12 cytokine family, is a potent proinflammatory cytokine produced by activated dendritic cells and macrophages. Cytokines in this family exist as heterodimers, and currently there are 4 known members: IL-12, IL-23, IL-27, and IL-35 (5). IL-23 is composed of the IL-12p40 subunit, which it shares with IL-12 and its own subunit, IL-23p19. IL-23 is known to stimulate immune responses and to drive differentiation of CD4+ cells into T helper (Th) type 17 cells. Although the role that IL-12 plays in type 1 diabetes (T1D) has been examined extensively (6–9), the role of IL-23 in diabetes, either directly or indirectly through induction of Th17 cells, is contradictory, depending on the conditions in which Th17 cells are generated. Several studies suggest Th17 cells play a significant role in the etiology of T1D, whereas others suggest Th17 cells are not involved (10–13). However, when Th17 cells do contribute to the etiology of T1D, it appears to rely on cells that can simultaneously produce IL-17 and IFN-γ (13–16). This current study was initially performed to examine whether increasing the expression of IL-23 by adenoviral gene transfer exacerbated or accelerated the establishment and progression of T1D in nonobese diabetic (NOD) mice. Surprisingly, prediabetic NOD mice injected with adenoviral vector encoding single-chain (sc)IL-23 (Ad.scIL-23) didn’t have accelerated onset of hyperglycemia. Instead, the treated mice developed visible symptoms of psoriasis, and histological analysis demonstrated the skin pathology of the classic features of psoriasis. Importantly, mice treated with Ad.scIL-23 also exhibited signs of psoriatic arthritis with evidence of intervertebral disc degeneration and synovial hyperplasia and erosion of the cartilage of the knee. Treatment with an anti–IL-17 antibody clearly implicated IL-17 in driving pathologies associated with psoriatic arthritis in joints, discs, and skin. These results demonstrated that systemic expression of scIL-23 in NOD mice is sufficient to drive not only psoriasis but also psoriatic arthritis. In addition, this novel mouse model of psoriatic arthritis can be used for analysis of disease progression and testing of novel therapeutics

Fisetin is a senotherapeutic that extends health and lifespanResearch in Context

Background: Senescence is a tumor suppressor mechanism activated in stressed cells to prevent replication of damaged DNA. Senescent cells have been demonstrated to play a causal role in driving aging and age-related diseases using genetic and pharmacologic approaches. We previously demonstrated that the combination of dasatinib and the flavonoid quercetin is a potent senolytic improving numerous age-related conditions including frailty, osteoporosis and cardiovascular disease. The goal of this study was to identify flavonoids with more potent senolytic activity. Methods: A panel of flavonoid polyphenols was screened for senolytic activity using senescent murine and human fibroblasts, driven by oxidative and genotoxic stress, respectively. The top senotherapeutic flavonoid was tested in mice modeling a progeroid syndrome carrying a p16INK4a-luciferase reporter and aged wild-type mice to determine the effects of fisetin on senescence markers, age-related histopathology, disease markers, health span and lifespan. Human adipose tissue explants were used to determine if results translated. Findings: Of the 10 flavonoids tested, fisetin was the most potent senolytic. Acute or intermittent treatment of progeroid and old mice with fisetin reduced senescence markers in multiple tissues, consistent with a hit-and-run senolytic mechanism. Fisetin reduced senescence in a subset of cells in murine and human adipose tissue, demonstrating cell-type specificity. Administration of fisetin to wild-type mice late in life restored tissue homeostasis, reduced age-related pathology, and extended median and maximum lifespan. Interpretation: The natural product fisetin has senotherapeutic activity in mice and in human tissues. Late life intervention was sufficient to yield a potent health benefit. These characteristics suggest the feasibility to translation to human clinical studies. Fund: NIH grants P01 AG043376 (PDR, LJN), U19 AG056278 (PDR, LJN, WLL), R24 AG047115 (WLL), R37 AG013925 (JLK), R21 AG047984 (JLK), P30 DK050456 (Adipocyte Subcore, JLK), a Glenn Foundation/American Federation for Aging Research (AFAR) BIG Award (JLK), Glenn/AFAR (LJN, CEB), the Ted Nash Long Life and Noaber Foundations (JLK), the Connor Group (JLK), Robert J. and Theresa W. Ryan (JLK), and a Minnesota Partnership Grant (AMAY-UMN#99)-P004610401–1 (JLK, EAA). Keywords: Senolytic, Aging, Progeria, Healthspan, Lifespan, Senescenc

Mesenchymal stem cell‐derived extracellular vesicles reduce senescence and extend health span in mouse models of aging

Aging drives progressive loss of the ability of tissues to recover from stress, partly through loss of somatic stem cell function and increased senescent burden. We demonstrate that bone marrow‐derived mesenchymal stem cells (BM‐MSCs) rapidly senescence and become dysfunctional in culture. Injection of BM‐MSCs from young mice prolonged life span and health span, and conditioned media (CM) from young BM‐MSCs rescued the function of aged stem cells and senescent fibroblasts. Extracellular vesicles (EVs) from young BM‐MSC CM extended life span of Ercc1−/− mice similarly to injection of young BM‐MSCs. Finally, treatment with EVs from MSCs generated from human ES cells reduced senescence in culture and in vivo, and improved health span. Thus, MSC EVs represent an effective and safe approach for conferring the therapeutic effects of adult stem cells, avoiding the risks of tumor development and donor cell rejection. These results demonstrate that MSC‐derived EVs are highly effective senotherapeutics, slowing the progression of aging, and diseases driven by cellular senescence. Extracellular vesicles from young bone marrow‐derived mesenchymal stem cells (MSC) reduce markers of senescence in vitro. EVs derived from MSCs generated from human embryonic stem cells reduced expression of senescence markers in culture and in vivo in accelerated and naturally aged mice and improved measures of healthspan. This work demonstrates the senotherapeutic potential of extracellular vesicles in suppressing senescence‐driven age related disease

Loss of DNA repair mechanisms in cardiac myocytes induce dilated cardiomyopathy.

Cardiomyopathy is a progressive disease of the myocardium leading to impaired contractility. Genotoxic cancer therapies are known to be potent drivers of cardiomyopathy, whereas causes of spontaneous disease remain unclear. To test the hypothesis that endogenous genotoxic stress contributes to cardiomyopathy, we deleted the DNA repair gene Ercc1 specifically in striated muscle using a floxed allele of Ercc1 and mice expressing Cre under control of the muscle-specific creatinine kinase (Ckmm) promoter or depleted systemically (Ercc1-/D mice). Ckmm-Cre+/- ;Ercc1-/fl mice expired suddenly of heart disease by 7 months of age. As young adults, the hearts of Ckmm-Cre+/- ;Ercc1-/fl mice were structurally and functionally normal, but by 6-months-of-age, there was significant ventricular dilation, wall thinning, interstitial fibrosis, and systolic dysfunction indicative of dilated cardiomyopathy. Cardiac tissue from the tissue-specific or systemic model showed increased apoptosis and cardiac myocytes from Ckmm-Cre+/- ;Ercc1-/fl mice were hypersensitive to genotoxins, resulting in apoptosis. p53 levels and target gene expression, including several antioxidants, were increased in cardiac tissue from Ckmm-Cre+/- ;Ercc1-/fl and Ercc1-/D mice. Despite this, cardiac tissue from older mutant mice showed evidence of increased oxidative stress. Genetic or pharmacologic inhibition of p53 attenuated apoptosis and improved disease markers. Similarly, overexpression of mitochondrial-targeted catalase improved disease markers. Together, these data support the conclusion that DNA damage produced endogenously can drive cardiac disease and does so mechanistically via chronic activation of p53 and increased oxidative stress, driving cardiac myocyte apoptosis, dilated cardiomyopathy, and sudden death