Attenuate Disease Progression Research Articles

CMS121 Partially Attenuates Disease Progression in Mouse Models of Huntington's Disease.

There are currently no drugs that meaningfully slow down the progression of Huntington's disease (HD). Moreover, drug candidates against a single molecular target have not had significant success. Therefore, a different approach to HD drug discovery is needed. Previously we showed that the flavonol fisetin is efficacious in mouse and fly models of HD (Hum. Mol. Gen. 20:261, 2011). It is also effective in animal models of Alzheimer's disease (AD), ischemic stroke, and the CNS complications of diabetes, all of which share some pathological features with HD. Potent derivatives of fisetin with improved pharmacology were made that maintain its multiple biological activities (J. Med. Chem. 55:378, 2012). From 160 synthetic fisetin derivatives, one, CMS121, was selected for further study in the context of HD based on pharmacological parameters and its efficacy in animal models of AD. Both R6/2 and YAC128 mouse models of HD were used in these studies. We examined motor function using multiple assays as well as survival. In the R6/2 mice, we also looked at the effects of CMS121 on striatal gene expression. In both models, we found a slowing of motor dysfunction and an increase in median life span. Interestingly, in the YAC128 mice, the effects on the slowing in motor function loss became increasingly more pronounced as the mice aged. CMS121 also reduced HD-driven changes in the expression of genes associated with the proteasome and oxidative phosphorylation. Overall, these results suggest that CMS121 could provide some benefits for HD patients, particularly with regard to increasing health span.

Gamma-secretase modulators: a promising route for the treatment of Alzheimer's disease.

Recent clinical data with three therapeutic anti-Aβ antibodies have demonstrated that removal of Aβ-amyloid plaques in early Alzheimer's disease (AD) can attenuate disease progression. This ground-breaking progress in AD medicine has validated both the amyloid cascade hypothesis and Aβ-amyloid as therapeutic targets. These results also strongly support therapeutic approaches that aim to reduce the production of amyloidogenic Aβ to prevent the formation of Aβ-pathology. One such strategy, so-called gamma-secretase modulators (GSM), has been thoroughly explored in preclinical settings but has yet to be fully tested in clinical trials. Recent scientific progress has shed new light on the role of Aβ in Alzheimer's disease and suggests that GSMs exhibit specific pharmacological features that hold great promise for the prevention and treatment of Alzheimer's disease. In this short review, we discuss the data that support why it is important to continue to progress in this class of compounds.

Increased thyroid hormones and autoantibodies levels associated with quasi-moyamoya disease

Objectives: The purpose of this study was to investigate whether elevated thyroid autoantibodies associated with quasi-Moyamoya Disease (MMD). Methods: We retrospectively investigated angiographically defined patients with quasi-MMD patients. We compared clinical features, serum thyroid autoantibody values and T cell levels between quasi-MMD patients and control subjects. Results: A total of 103 patients with MMD and an equal number of healthy control subjects (n=103) were included. The prevalence of elevated thyroid autoantibodies (P=0.008) were significantly higher in patients with MMD than in control subjects. The increased thyroid hormone (OR, 11.03; 95% CI, 1.22 to 96.13; P=0.034) and Increased thyroid autoantibodies (OR, 6.38; 95% CI, 1.12 to 46.52; P=0.040) were significantly associated with increased risks of quasi-MMD, respectively. The antithyroid therapy can significantly attenuate disease progression than without special treatment in quasi-MMD patients with thyroid disease (P=0.041).

Six-month effects of supervised exercise on walking ability and health-related factors in peripheral arterial disease: a pilot study.

In peripheral arterial disease (PAD) patients with intermittent claudication (IC), the combination of aerobic and resistance exercises could counteract muscle loss and attenuate disease progression. This study analyzed the effects of six months of a combined exercise program on walking ability, lower limb body composition, cardiovascular risk factors, and Ankle-Brachial Index (ABI). Twenty-three patients (age 63.2±1.5 years and ABI 0.58±0.07) with PAD and IC were allocated to a control group (CG) or a supervised exercise group (SUP). Ten patients underwent six months of treadmill walking combined with resistance exercises, three times a week. The CG (N.=13) received a recommendation for walking. All patients were measured at baseline (M0), after three months (M3), and six months (M6). During constant treadmill protocol, the claudication onset time/distance (COT/COD), absolute claudication time/distance (ACT/ACD), and number of pauses of overall patients significantly improved at M3 and M6. Between groups were found significant differences in COT and COD at M6 (P=0.005 and P=0.007, respectively); and in ACT and ACD at M3 (P=0.003 for both) and at M6 (P=0.005 and P=0.005, respectively), with major improvements in the SUP. Over the six months, a significant group effect was found in fat-free mass (P=0.041) and predicted muscle mass (P=0.039) of the lower ABI leg, with greater improvements in the SUP. A supervised exercise program that combines aerobic and resistance training improves PAD symptoms and has additional benefits for patients. Patients in the program showed improvements in walking ability, lower-limb body composition, perceived exertion, and heart rate during treadmill walking.

Updates on Emerging Interventions for Autosomal Recessive ABCA4-Associated Stargardt Disease.

Autosomal recessive Stargardt disease (STGD1) is an inherited retinal degenerative disease associated with a mutated ATP-binding cassette, subfamily A, member 4 (ABCA4) gene. STGD1 is the most common form of juvenile macular degeneration with onset in late childhood to early or middle adulthood and causes progressive, irreversible visual impairment and blindness. No effective treatment is currently available. In the present article, we review the most recent updates in clinical trials targeting the management of STGD1, including gene therapy, small molecule therapy, and stem cell therapy. In gene therapy, dual adeno-associated virus and non-viral vectors have been successful in delivering the human ABCA4 gene in preclinical studies. For pharmaceutical therapies ALK-001, deuterated vitamin A shows promise with preliminary data for phase 2 trial, demonstrating a decreased atrophy growth rate after two years. Stem cell therapy using human pluripotent stem cell-derived retinal pigment epithelium cells demonstrated long-term safety three years after implantation and visual acuity improvements in the first two years after initiation of therapy. Many other treatment options have ongoing investigations and clinical trials. While multiple potential interventions have shown promise in attenuating disease progression, further exploration is necessary to demonstrate treatment safety and efficacy.

Brensocatib (an oral, reversible inhibitor of dipeptidyl peptidase-1) attenuates disease progression in two animal models of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a painful and incurable disease characterized by chronic joint inflammation and a progressive destruction of cartilage and bone. Although current treatments have improved clinical outcomes for some patients, the high relapse rates and sizeable proportion of non-responders emphasize the need for further research. Arthritic joints are massively infiltrated by neutrophils, which influence inflammatory and immune processes by releasing cytokines, chemokines, eicosanoids, and neutrophil serine proteases (NSPs) - all of which are known to contribute to RA initiation and progression. Active NSPs are generated from zymogens at the promyelocytic stage of neutrophil differentiation under the action of dipeptidyl peptidase 1 (DPP-1) and DPP-1 knockout mice are resistant to the development of arthritis. Thus, DPP-1 inhibition represents a promising therapeutic approach in RA. In this study, we assessed the efficacy of a potent and highly selective DPP-1 inhibitor, brensocatib, in two well established RA models - rat collagen-induced arthritis (CIA) and mouse collagen antibody-induced arthritis (CAIA). In both models, brensocatib at 3 and 30 mg/kg/day significantly reduced bone marrow NSP levels, in keeping with prior pharmacodynamic studies in rodents. More importantly, brensocatib treatment significantly improved disease score at both dosages in both rodent models. In the mouse CAIA model, brensocatib even proved at least as potent as anti-TNF antibodies in diminishing both the histopathological score and neutrophil infiltration into arthritic joints. Together, these results show that brensocatib alters RA disease progression in rodents and supports the need for its further evaluation as a potential therapeutic option, or to complement existing RA treatments.

COMPARISON OF CONVENTIONAL DISEASE MODIFYING ANTI-RHEUMATIC DRUGS WITH AND WITHOUT INTRA-ARTICULAR PLATELET RICH PLASMA INJECTION IN THE ALTERATION OF DISEASE PROGRESSION OF EARLY RHEUMATOID ARTHRITIS: A RANDOMIZED CONTROLLED TRIAL

Background and Objectives:Rheumatoid arthritis (RA) is a symmetric inflammatory polyarticular arthritis, where autoimmune response causes chronic inflammation and destruction of joints leading to deformities and affecting the activities of daily living. ACR recommends early diagnosis and aggressive treatment of RA with combination disease modifying anti-rheumatic drugs (DMARDs). Although it attenuates disease progression, long-term remissions have not been adequately achieved. Injection of platelet rich plasma(PRP) decrease joint inflammation by modulating synovial cell proliferation, differentiation and inhibition of catabolic pathways. Aim:to compare conventional DMARDs with and without intra-articular platelet rich plasma injection in wrist joint in the alteration of disease progression of early rheumatoid arthritis. Methods:A randomized controlled trial, done among newly diagnosed patients of Rheumatoid Arthritis affecting wrist joint, in Department of PMR, RIMS. Data collected in pretested structured proforma and analysis for Pain, Disease activity, USG grading and hand function done using Chi-square test, students t-test, repeated measure ANOVA test. Results:There is significant improvement in VAS, DAS28, Cochin scale and USG Grade in study group at the end of 8 weeks, however at 20 weeks it is found to be comparable and p-value is not significant. Conclusion:Addition of Intra articular PRP in wrist along with DMARDs has early and better improvement in pain and delay disease progression. Intra-articular PRP injection under USG guidance helps in reduction of synovial inflammation and pain, improvement in function and a safe treatment modality. Early treatment and reduction of disease activity by intervention in targeted joint may delay joint deformity and disability.

Caspofungin-induced β(1,3)-glucan exposure in Candida albicans is driven by increased chitin levels.

To successfully induce disease, Candida albicans must effectively evade the host immune system. One mechanism used by C. albicans to achieve this is to mask immunogenic β(1,3)-glucan epitopes within its cell wall under an outer layer of mannosylated glycoproteins. Consequently, induction of β(1,3)-glucan exposure (unmasking) via genetic or chemical manipulation increases fungal recognition by host immune cells in vitro and attenuates disease during systemic infection in mice. Treatment with the echinocandin caspofungin is one of the most potent drivers of β(1,3)-glucan exposure. Several reports using murine infection models suggest a role for the immune system, and specifically host β(1,3)-glucan receptors, in mediating the efficacy of echinocandin treatment in vivo. However, the mechanism by which caspofungin-induced unmasking occurs is not well understood. In this report, we show that foci of unmasking co-localize with areas of increased chitin within the yeast cell wall in response to caspofungin, and that inhibition of chitin synthesis via nikkomycin Z attenuates caspofungin-induced β(1,3)-glucan exposure. Furthermore, we find that both the calcineurin and Mkc1 mitogen-activated protein kinase pathways work synergistically to regulate β(1,3)-glucan exposure and chitin synthesis in response to drug treatment. When either of these pathways are interrupted, it results in a bimodal population of cells containing either high or low chitin content. Importantly, increased unmasking correlates with increased chitin content within these cells. Microscopy further indicates that caspofungin-induced unmasking correlates with actively growing cells. Collectively, our work presents a model in which chitin synthesis induces unmasking within the cell wall in response to caspofungin in growing cells. IMPORTANCE Systemic candidiasis has reported mortality rates ranging from 20% to 40%. The echinocandins, including caspofungin, are first-line antifungals used to treat systemic candidiasis. However, studies in mice have shown that echinocandin efficacy relies on both its cidal impacts on Candida albicans, as well as a functional immune system to successfully clear invading fungi. In addition to direct C. albicans killing, caspofungin increases exposure (unmasking) of immunogenic β(1,3)-glucan moieties. To evade immune detection, β(1,3)-glucan is normally masked within the C. albicans cell wall. Consequently, unmasked β(1,3)-glucan renders these cells more visible to the host immune system and attenuates disease progression. Therefore, discovery of how caspofungin-induced unmasking occurs is needed to elucidate how the drug facilitates host immune system-mediated clearance in vivo. We report a strong and consistent correlation between chitin deposition and unmasking in response to caspofungin and propose a model in which altered chitin synthesis drives increased unmasking during drug exposure.

Brensocatib, an oral, reversible inhibitor of dipeptidyl peptidase 1, mitigates interferon-α-accelerated lupus nephritis in mice.

Neutrophils have been implicated in initiating and perpetuating systemic lupus erythematosus and the resultant kidney damage in lupus nephritis (LN) patients, in part through an excessive release of neutrophil serine proteases (NSPs). NSP zymogens are activated by dipeptidyl peptidase 1 (DPP1) during neutrophil maturation and released by mature neutrophils in response to inflammatory stimuli. Thus, a potential strategy to attenuate disease progression in LN would be to inhibit DPP1. We tested whether brensocatib, a highly selective and reversible DPP1 inhibitor, could mitigate LN progression in an interferon-alpha (IFNα)-accelerated NZB/W F1 mouse model. To confirm brensocatib's pharmacodynamic effect on NSPs in this mouse strain, repeated dose studies were conducted for 7 and 14 days in naïve NZB/W F1 mice via oral gavage twice a day. Brensocatib at 2 and 20 mg/kg/day achieved a significant reduction in bone marrow NSP activities after 7 days of daily administration. To initiate LN disease progression, the mice were injected with an IFNα-expressing adenovirus. After 2 weeks, three brensocatib doses (or vehicle) were administered for 6 more weeks. Throughout the 8-week study, brensocatib treatment (20 mg/kg/day) significantly reduced the occurrence of severe proteinuria compared to the vehicle control. Brensocatib treatment also entailed a significant reduction in the urine albumin-to-creatinine ratio, indicating decreased kidney damage, as well as a significant reduction in blood urea nitrogen level, suggesting improved renal function. Based on kidney histopathology analysis, brensocatib treatment significantly lowered both the renal tubular protein score and the nephropathy score compared to the vehicle group. A trend towards reduced glomerulonephritis score with brensocatib treatment was also observed. Lastly, brensocatib significantly reduced LN mouse kidney infiltration by various inflammatory cells. In conclusion, these results suggest that brensocatib alters disease progression in LN mice and warrant further evaluation of DPP1 inhibition in LN.

#4420 EARLY DAPAGLIFLOZIN UTILIZATION FOR CHRONIC KIDNEY DISEASE TREATMENT IN JAPAN

Abstract Background and Aims In 2021, dapagliflozin was approved for the treatment of adults with chronic kidney disease (CKD) in Japan. This observational analysis is part of the multinational OPTIMISE-CKD study program and aims to describe early dapagliflozin utilization in CKD following approval in Japan, contextualized by contemporary CKD management. Method Adult patients with CKD in hospital claims data from the Medical Data Vision Co. Ltd database (MDV; Tokyo, Japan) during 1 March 2020 – 28 February 2022 were included. CKD was defined as any of the following: 2 estimated glomerular filtration rate (eGFR) measures ≥90 days apart of which the second was ≤75 ml/min/1.73 m2, or a CKD diagnosis code. Patients with type 1 diabetes, gestational diabetes, <365 days continuous enrolment or who were on dialysis were excluded. The remaining patients were considered eligible for dapagliflozin 10 mg treatment. Dapagliflozin initiators had a recorded prescription for dapagliflozin 10 mg on or after 26 August 2021, and no previous dapagliflozin 10 mg prescription. Results A total of 2,297 patients in MDV met the study criteria and had initiated dapagliflozin during the first 6 months since approval. The median age was 70 years (interquartile range; IQR 57-79 years), 33% were female and 50% had a recorded CKD diagnosis. For the 594 initiators who had eGFR data, the CKD stage distribution was as follows: 27% stage 1-2, 28% stage 3a, 28% stage 3b, 16% stage 4 and 1% stage 5 (non-dialysis). The average eGFR among these patients was 49 ml/min/1.73 m2 (standard deviation; SD 19 ml/min/1.73 m2). The most common comorbidities were heart failure (37%), hypertension (34%) and type 2 diabetes (34%). Renin-angiotensin system inhibitors (RASi; 58%), calcium channel blockers (37%) and lipid-lowering drugs (37%) were commonly prescribed. Dapagliflozin 10 mg was most commonly initiated in nephrology departments (42%), with a mean initial prescription duration of 41 days (SD 22.6 days). The 180,462 patients who did not initiate dapagliflozin 10 mg were older on average (median 78 years; IQR 69-85 years), and 42% were female. The comorbidity profiles were similar, except that 24% of patients had type 2 diabetes. The distribution of CKD stages among the 95,465 patients with available data was slightly different: 59% stage 1-2, 22% stage 3a, 11% stage 3b, 6% stage 4 and 3% stage 5 (non-dialysis). Only 23% of patients were prescribed RASi and 41% were prescribed any kind of cardiovascular treatment. Conclusion Early use of dapagliflozin in Japan was observed in a broad range of patients with CKD with respect to baseline characteristics. These preliminary data suggest that there is a large proportion of CKD patients who may be eligible, but not yet treated with dapagliflozin 10 mg, which has shown to attenuate disease progression and reduce adverse outcomes. Considering that the majority of prescriptions was made in nephrology departments, there might be opportunities in other healthcare specialties to improve CKD diagnosis and treatment. Additional analyses, including sensitivity analyses using more conservative CKD definitions, could further specify the unmet need and highlight activities that may increase prescriptions among patients who might benefit from treatment. Further analyses across more patients and countries in the OPTIMISE-CKD program will provide additional insight into the real-world experience of novel treatments for CKD and facilitate optimized CKD management.

Innate defenses to intestinal cell death in necrotizing enterocolitis – spotlight on macrophage efferocytosis and its efficacy in rescuing inflamed intestinal mucosa

Introduction and aim. Necrotizing enterocolitis (NEC) is a grave gastrointestinal disease of preterm infants which is widely prevalent in the neonatal intensive care units. Current treatment options are very limited with high mortality and morbidity. With no disease specific interventions, understanding nascent cellular events that occur immediately after microbial insult can offer insights for devising novel treatment options for curtailing the disease progression in NEC. In this regard, intestinal cell death in NEC is a primordial cell-signaling event and is regarded as a harbinger of future pathological derangements such as increased intestinal permeability, intestinal dys-homeostasis, and systemic inflammation. Material and methods. We performed PubMed search of relevant articles that describes the host response to intestinal cell death in NEC by cellular battalion including dendritic cells, lymphocytes, neutrophils and macrophages which are important in containing intestinal inflammation. Analysis of the literature. We particularly focused this review on enumerating macrophage efferocytosis, and pertinent novel treatment modalities based on this physiological process that has inherent capability for down regulating inflammation and promoting tissue repair in NEC. We highlighted its mechanistic aspect including mediators, receptors and signaling mechanisms and its physiological significance. Conclusion. Macrophage efferocytosis is an overlooked and undervalued physiological defense mechanism to clear the dying intestinal epithelial cells for facilitating tissue healing and restoring the intestinal homeostasis. Any impairment of this critical defense mechanism can result in rapid clinical progression and systemic complications. Understanding its importance in the pathogenesis of NEC is important for designing novel therapeutic interventions to attenuate disease progression.

PO-01-147 ACTIVATION OF AUTOPHAGY ATTENUATES PROGRESSION OF ARRHYTHMOGENIC RIGHT VENTRICULAR CARDIOMYOPATHY

Studies of autophagy in ARVC are quite rare. Even autophagy had the opposite effects in desmin-related cardiomyopathy and dilated cardiomyopathy. This study aimed to explore the regulatory role of autophagy in the progression of ARVC. The ARVC cardiomyocytes (HL-1/shJup) were treated with autophagy activator (rapamycin) and inhibitor (chloroquine) to manipulate autophagy activity. Western blotting and immunofluorescence determined the changes of disease progression and signaling pathways. Then, autophagy was manipulated in cardiac tissue-specific Jup knockout mice to confirm the cell-derived findings. The autophagy activity of HL-1/shJup cell line was higher than that of control cells. In HL-1/shJup cells, the upstream signaling pathway of autophagy was active. During rapamycin treatment, the proteins related to lipogenesis and fibrogenesis in HL-1/shJup cells were reduced. In contrast, chloroquine treatment had the opposite effect. Moreover, rapamycin decreased and chloroquine increased intracellular oil droplet accumulation in HL-1/shJup cells. In addition, there was no significant change in Wnt/Hippo signaling pathways in cardiomyocytes after rapamycin treatment. Instead, the protein expression of the transcription factor SREBP1 that regulates lipogenesis and lysyl oxidase (LOX) that promotes fibrosis were related to the regulatory mechanism of autophagy. In ARVC mice, activated/inhibited autophagy also reduced/increased RV distance, arrhythmia, and lipogenesis through SREBP1, but not in wild-type mice. Autophagy plays a protective role in ARVC by regulating lipogenesis to attenuate disease progression. Based on this study, autophagy activators may be used in combination with existing drugs in patients with ARVC disease in the future to provide additional protection for early heart failure or heart transplantation.

A new immunodeficient Duchenne muscular dystrophy rat model to evaluate engraftment after human cell transplantation.

Duchenne muscular dystrophy (DMD) is an X-linked fatal muscular disease, affecting one in 3,500 live male births worldwide. Currently, there is no cure for this disease, except for steroid-based treatment to attenuate disease progression. Cell transplantation therapy is a promising therapeutic approach, however, there is a lack of appropriate animal models to conduct large-scale preclinical studies using human cells, including biochemical and functional tests. Here, we established an immunodeficient DMD rat model and performed exhaustive pathological analysis and transplantation efficiency evaluation to assess its suitability to study DMD. Our DMD rat model exhibited histopathological characteristics similar to those observed in human patients with DMD. Human myoblasts demonstrated successful engraftment following transplantation into these rats. Therefore, this immunodeficient DMD rat model would be useful in preclinical studies to develop cellular transplantation therapies for DMD.

Abstract 6225: SERPINB3 promotes the aggressive basal-like/squamous subtype and correlates with poor prognosis in pancreatic ductal adenocarcinoma through metabolic reprogramming

Abstract In this study, we aimed to find genes with a key function in the development of molecular subtypes in pancreatic ductal adenocarcinoma (PDAC). Through transcriptome analysis, we discovered that the endogenous serine/cysteine proteinase inhibitor SERPINB3 (squamous cell carcinoma antigen 1, SCCA1) was distinctively upregulated in the basal-like/squamous subtype, known as an aggressive subtype, and this upregulation associated with decreased patient survival in both a test (N=123) and a validation cohort (N=84). In additional investigations of the tumor metabolome and transcriptome using PDAC patient tumors and cell lines, SERPINB3 and the basal-like/squamous subtype showed a robust relationship with upregulated levels of amino acids (e.g., hydroxyproline) whereas SERPINB3 promoted a gene signature indicative of the basal-like/squamous subtype. Additional mechanistic studies revealed that SERPINB3 and hydroxyproline promoted the migration/invasion of PDAC cells. Moreover, SERPINB3 also promoted metastasis in an orthotopic mouse model of PDAC through stromal factors that increased tumor microvessel density. To conclude, inhibiting SERPINB3 function may attenuate disease progression of the basal-like/squamous subtype in PDAC through changes to the tumor stroma and tumor metabolism. Citation Format: Yuuki Ohara, Wei Tang, Liu Huaitian, Shouhui Yang, Peijun He, Helen Cawley, Paloma Valenzuela, Lin Zhang, Jochen Gaedcke, B. Michael Ghadimi, Matthias M. Gaida, Frank Bergmann, H. Richard Alexander, Nader Hanna, Stefan Ambs, S. Perwez Hussain. SERPINB3 promotes the aggressive basal-like/squamous subtype and correlates with poor prognosis in pancreatic ductal adenocarcinoma through metabolic reprogramming [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6225.

Ketones and the Heart: Metabolic Principles and Therapeutic Implications.

The ketone bodies beta-hydroxybutyrate and acetoacetate are hepatically produced metabolites catabolized in extrahepatic organs. Ketone bodies are a critical cardiac fuel and have diverse roles in the regulation of cellular processes such as metabolism, inflammation, and cellular crosstalk in multiple organs that mediate disease. This review focuses on the role of cardiac ketone metabolism in health and disease with an emphasis on the therapeutic potential of ketosis as a treatment for heart failure (HF). Cardiac metabolic reprogramming, characterized by diminished mitochondrial oxidative metabolism, contributes to cardiac dysfunction and pathologic remodeling during the development of HF. Growing evidence supports an adaptive role for ketone metabolism in HF to promote normal cardiac function and attenuate disease progression. Enhanced cardiac ketone utilization during HF is mediated by increased availability due to systemic ketosis and a cardiac autonomous upregulation of ketolytic enzymes. Therapeutic strategies designed to restore high-capacity fuel metabolism in the heart show promise to address fuel metabolic deficits that underpin the progression of HF. However, the mechanisms involved in the beneficial effects of ketone bodies in HF have yet to be defined and represent important future lines of inquiry. In addition to use as an energy substrate for cardiac mitochondrial oxidation, ketone bodies modulate myocardial utilization of glucose and fatty acids, two vital energy substrates that regulate cardiac function and hypertrophy. The salutary effects of ketone bodies during HF may also include extra-cardiac roles in modulating immune responses, reducing fibrosis, and promoting angiogenesis and vasodilation. Additional pleotropic signaling properties of beta-hydroxybutyrate and AcAc are discussed including epigenetic regulation and protection against oxidative stress. Evidence for the benefit and feasibility of therapeutic ketosis is examined in preclinical and clinical studies. Finally, ongoing clinical trials are reviewed for perspective on translation of ketone therapeutics for the treatment of HF.

Bruton's tyrosine kinase inhibition attenuates disease progression by reducing renal immune cell invasion in mice with hemolytic-uremic syndrome.

Hemolytic-uremic syndrome (HUS) can occur as a complication of an infection with Shiga-toxin (Stx)-producing Escherichia coli. Patients typically present with acute kidney injury, microangiopathic hemolytic anemia and thrombocytopenia. There is evidence that Stx-induced renal damage propagates a pro-inflammatory response. To date, therapy is limited to organ-supportive strategies. Bruton's tyrosine kinase (BTK) plays a pivotal role in recruitment and function of immune cells and its inhibition was recently shown to improve renal function in experimental sepsis and lupus nephritis. We hypothesized that attenuating the evoked immune response by BTK-inhibitors (BTKi) ameliorates outcome in HUS. We investigated the effect of daily oral administration of the BTKi ibrutinib (30 mg/kg) and acalabrutinib (3 mg/kg) in mice with Stx-induced HUS at day 7. After BTKi administration, we observed attenuated disease progression in mice with HUS. These findings were associated with less BTK and downstream phospholipase-C-gamma-2 activation in the spleen and, subsequently, a reduced renal invasion of BTK-positive cells including neutrophils. Only ibrutinib treatment diminished renal invasion of macrophages, improved acute kidney injury and dysfunction (plasma levels of NGAL and urea) and reduced hemolysis (plasma levels of bilirubin and LDH activity). In conclusion, we report here for the first time that BTK inhibition attenuates the course of disease in murine HUS. We suggest that the observed reduction of renal immune cell invasion contributes - at least in part - to this effect. Further translational studies are needed to evaluate BTK as a potential target for HUS therapy to overcome currently limited treatment options.

Efficacy and Safety Evaluation of Mometasone Furoate in Treating Ocular Inflammation.

Mometasone furoate (MF) is a medium-potency synthetic glucocorticosteroid with anti-inflammatory, antipruritic, and vasoconstrictive properties. However, its role in the treatment of ocular inflammation has not yet been explored. This work investigated the anti-inflammatory activity of MF in ocular tissues. First, the in vivo safety of the intravitreal (IVT) injection of MF (80, 160, and 240 µg) was evaluated via clinical examination (including the assessment of intraocular pressure), electroretinography (ERG), and histopathology. Second, MF was tested in an experimental model of bacillus Calmette-Guérin (BCG)-induced uveitis in Wistar rats. Intraocular inflammation was then evaluated via a slit-lamp and fundus examination, ERG, histopathology, and the quantification of pro-inflammatory markers. Intravitreal MF showed no toxicity in all the investigated doses, with 160 µg leading to attenuated disease progression and improvement in clinical, morphological, and functional parameters. There was a significant reduction in the levels of inflammatory markers (myeloperoxidase, interleukins 6 and 1β, CXCL-1, and tumor necrosis factor-alpha) when compared to the levels in untreated animals. Therefore, MF should be further investigated as a promising drug for the treatment of ocular inflammation.

Olfactory Ecto-mesenchymal Stem Cell-derived Exosomes Ameliorate Murine Sjögren's Syndrome via Suppressing Tfh Cell Response.

To investigate the effect of olfactory ecto-mesenchymal stem cell-derived exosomes (OE-MSC-Exos) on T follicular helper (Tfh) cell response and their implication in treating experimental Sjögrens syndrome (ESS). C57BL/6 mice were immunized with salivary glands (SG) proteins to induce ESS mouse model. OE-MSC-Exos were added to the Tfh cell polarization condition, and the proportion of Tfh cells was detected by FCM. The PD-L1 of OE-MSCs was silenced with small interfering RNA to extract siPD-L1-OE-MSC-Exos. We found that transfer of OE-MSC-Exos markedly attenuated disease progression and reduced Tfh cell response in mice with ESS. In culture, OE-MSC-Exos potently inhibited the differentiation of Tfh cells from naïve T cells. Moreover, OE-MSC-Exos expressed high level of the ligand for the programmed cell death protein 1 (PD-L1), knocking down PD-L1 expression in OE-MSC-Exos significantly decreased their capacity to suppress Tfh cell differentiation in vitro. Consistently, transfer of OE-MSC-Exos with PD-L1 knockdown exhibited profoundly diminished therapeutic effect in ESS mice, accompanied with sustained Tfh cell response and high levels of autoantibody production. Our results suggest that OE-MSC-Exos may exert their therapeutic effect in ameliorating ESS progression via suppressing Tfh cell response in a PD-L1-dependent manner.

Dysregulation of HNF1B/Clusterin axis enhances disease progression in a highly aggressive subset of pancreatic cancer patients.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy and is largely refractory to available treatments. Identifying key pathways associated with disease aggressiveness and therapeutic resistance may characterize candidate targets to improve patient outcomes. We used a strategy of examining the tumors from a subset of PDAC patient cohorts with the worst survival to understand the underlying mechanisms of aggressive disease progression and to identify candidate molecular targets with potential therapeutic significance. Non-negative matrix factorization (NMF) clustering, using gene expression profile, revealed three patient subsets. A 142-gene signature specific to the subset with the worst patient survival, predicted prognosis and stratified patients with significantly different survival in the test and validation cohorts. Gene-network and pathway analysis of the 142-gene signature revealed dysregulation of Clusterin (CLU) in the most aggressive patient subset in our patient cohort. Hepatocyte nuclear factor 1 b (HNF1B) positively regulated CLU, and a lower expression of HNF1B and CLU was associated with poor patient survival. Mechanistic and functional analyses revealed that CLU inhibits proliferation, 3D spheroid growth, invasiveness and epithelial-to-mesenchymal transition (EMT) in pancreatic cancer cell lines. Mechanistically, CLU enhanced proteasomal degradation of EMT-regulator, ZEB1. In addition, orthotopic transplant of CLU-expressing pancreatic cancer cells reduced tumor growth in mice. Furthermore, CLU enhanced sensitivity of pancreatic cancer cells representing aggressive patient subset, to the chemotherapeutic drug gemcitabine. Taken together, HNF1B/CLU axis negatively regulates pancreatic cancer progression and may potentially be useful in designing novel strategies to attenuate disease progression in PDAC patients.

MOLECULAR DOCKING STUDIES OF MONOMERIC WILDTYPE AND MUTANT (H81A, H49R) SOD1 WITH EDARAVONE AND RILUZOLE

Objective: Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder that causes progressive loss of the upper and lower motor neurons in the brain and spinal cord. SOD1 was the first gene linked to ALS, in which more than 150 mutations throughout the sequence of the protein have been found to be associated with ALS. Methods: The drugs that can interact and inhibit the misfolding or revert the misconformation of the protein can be useful in the treatment of ALS. Monomer apo SOD1-WT and MBR SOD1 mutants (H81A, H49R) were docked with the only two FDA-approved drugs for ALS that are edaravone and riluzole to assess if the ALS patients carrying these particular protein aggregates will derive any therapeutic efficacy. Results: The drugs were found to interact with both wild-type and mutant SOD1 at different positions and the type of interaction, degree of interaction and their binding energies were determined. SOD1-WT has hydrophobic interactions at V103, H110, and R115 and hydrogen bond at H110 with edaravone. SOD1-WT has hydrophobic interactions at T54, F64 and hydrogen bond at D52, T58 with riluzole. Both SOD1 mutants has a hydrogen bond at its H46 residue for both drugs. SOD1-H49R mutant has hydrophobic interactions at F45 with riluzole. SOD1-H81A mutant has hydrophobic interactions at F45 with edaravone. Conclusion: Both the mutants L42A residue has a hydrophobic interaction with edaravone. Interaction of drugs with protein and its mutants may make it possible to restore the stability of SOD1 structure and attenuate disease progression in ALS patients carrying these mutations.