Molecular Targets to Alleviate Enteric Neuropathy and Gastrointestinal Dysfunction.

Enteric neuropathy underlies long-term gastrointestinal (GI) dysfunction associated with several pathological conditions. Our previous studies have demonstrated that structural and functional changes in the enteric nervous system (ENS) result in persistent alterations of intestinal functions long after the acute insult. These changes lead to aberrant immune response and chronic dysregulation of the epithelial barrier. Damage to the ENS is prognostic of disease progression and plays an important role in the recurrence of clinical manifestations. This suggests that the ENS is a viable therapeutic target to alleviate chronic intestinal dysfunction. Our recent studies in preclinical animal models have progressed into the development of novel therapeutic strategies for the treatment of enteric neuropathy in various chronic GI disorders. We have tested the anti-inflammatory and neuroprotective efficacy of novel compounds targeting specific molecular pathways. Ex vivo studies in human tissues freshly collected after resection surgeries provide an understanding of the molecular mechanisms involved in enteric neuropathy. In vivo treatments in animal models provide data on the efficacy and the mechanisms of actions of the novel compounds and their combinations with clinically used therapies. These novel findings provide avenues for the development of safe, cost-effective, and highly efficacious treatments of GI disorders.

Divergent Adaptations in Autonomic Nerve Activity and Neuroimmune Signaling Associated With the Severity of Inflammation in Chronic Colitis.

BACKGROUND: The autonomic nervous system (ANS) is thought to play a critical role in the anti-inflammatory reflex pathway in acute colitis via its interaction with the spleen and colon. Inflammation in the intestine is associated with a blunting of vagal signaling and increased sympathetic activity. As a corollary, methods to restore sympatho-vagal balance are being investigated as therapeutic strategies for the treatment of intestinal inflammation. Nevertheless, it is indefinite whether these autonomic signaling adaptations in colitis are detrimental or beneficial to controlling intestinal inflammation. In this study, models of moderate and severe chronic colitis are utilized to resolve the correlations between sympatho-vagal signaling and the severity of intestinal inflammation. METHODS: Spleens and colons were collected from Winnie (moderate colitis), Winnie-Prolapse (severe colitis), and control C57BL/6 mice. Changes to the size and histomorphology of spleens were evaluated. Flow cytometry was used to determine the expression of adrenergic and cholinergic signaling proteins in splenic B and T lymphocytes. The inflammatory profile of the spleen and colon was determined using a RT-PCR gene array. Blood pressure, heart rate, splanchnic sympathetic nerve and vagus nerve activity were recorded. RESULTS: Spleens and colons from Winnie and Winnie-Prolapse mice exhibited gross abnormalities by histopathology. Genes associated with a pro-inflammatory response were upregulated in the colons from Winnie and further augmented in colons from Winnie-Prolapse mice. Conversely, many pro-inflammatory markers were downregulated in the spleens from Winnie-Prolapse mice. Heightened activity of the splanchnic nerve was observed in Winnie but not Winnie-Prolapse mice. Conversely, vagal nerve activity was greater in Winnie-Prolapse mice compared with Winnie mice. Splenic lymphocytes expressing α1 and ß2 adrenoreceptors were reduced, but those expressing α7 nAChR and producing acetylcholine were increased in Winnie and Winnie-Prolapse mice. CONCLUSIONS: Sympathetic activity may correlate with an adaptive mechanism to reduce the severity of chronic colitis. The Winnie and Winnie-Prolapse mouse models of moderate and severe chronic colitis are well suited to examine the pathophysiology of progressive chronic intestinal inflammation.

In this study we use mouse models of moderate and severe colitis to resolve the relationship between autonomic and neuroimmune signaling with inflammation. Increased expression of cholinergic markers on immune cells correlated with an anti-inflammatory profile in the spleen, consistent with activation of the splenic cholinergic anti-inflammatory pathway in mice with spontaneous chronic colitis. However, enhanced sympathetic signaling occurred in mice with a less severe phenotype of colitis, which could represent an adaptive mechanism to mitigate the progression of intestinal inflammation.

The comparative anti-cancer effects of krill oil and oxaliplatin in an orthotopic mouse model of colorectal cancer.

BACKGROUND: Our in vitro studies demonstrated that krill oil (KO) has anti-cancer potential. This study aimed to compare the anti-cancer effects of KO with a commonly used chemotherapeutic drug, oxaliplatin and to identify the molecular mechanisms associated with KO supplementation in a mouse model of colorectal cancer (CRC). METHODS: Thirty-six male Balb/c mice were randomly divided into six groups. Five groups received standard chow diet supplemented with KO (150 g/kg)), corn oil (150 g/kg), KO combined with ½ dose of oxaliplatin (1.5 mg/kg body weight/3 times per week), corn oil combined with ½ dose of oxaliplatin (1.5 mg/kg body weight/3 times per week), or a full dose of oxaliplatin (3 mg/kg body weight/3 times per week). The control (sham) group received a standard chow diet. Treatments started three weeks before and continued for three weeks after orthotopic CRC induction. The number of metastases, tumour weight and volume were quantified ex-vivo. The expression of cytochrome c, cleaved caspase-9 and -3, DNA damage, PD-L1, PD-L2 and HSP-70 were determined. RESULTS: A significant reductions in the weight and volume of tumours were observed in mice treated with KO and KO plus a ½ dose of oxaliplatin compared to the sham group, similar to oxaliplatin-treated mice. KO, and KO plus ½ dose of oxaliplatin significantly increased the expression of cytochrome c, cleaved caspase-9 and -3, and DNA damage and decreased expression of PD-L1, PD-L2 and HSP-70 in tumour tissues compared to the sham group. CONCLUSIONS: The in vivo anti-cancer effects of KO are comparable with oxaliplatin. Thus, dietary KO supplementation has a great potential as a therapeutic/adjunctive agent for CRC treatment.

Characterization of Skeletal Phenotype and Associated Mechanisms With Chronic Intestinal Inflammation in the Winnie Mouse Model of Spontaneous Chronic Colitis.

BACKGROUND: Osteoporosis is a common extraintestinal manifestation of inflammatory bowel disease (IBD). However, studies have been scarce, mainly because of the lack of an appropriate animal model of colitis-associated bone loss. In this study, we aimed to decipher skeletal manifestations in the Winnie mouse model of spontaneous chronic colitis, which carries a MUC2 gene mutation and closely replicates ulcerative colitis. In our study, Winnie mice, prior to the colitis onset at 6 weeks old and progression at 14 and 24 weeks old, were compared with age-matched C57BL/6 controls. We studied several possible mechanisms involved in colitis-associated bone loss. METHODS: We assessed for bone quality (eg, microcomputed tomography [micro-CT], static and dynamic histomorphometry, 3-point bending, and ex vivo bone marrow analysis) and associated mechanisms (eg, electrochemical recordings for gut-derived serotonin levels, real-time polymerase chain reaction [qRT-PCR], double immunofluorescence microscopy, intestinal inflammation levels by lipocalin-2 assay, serum levels of calcium, phosphorus, and vitamin D) from Winnie (6-24 weeks) and age-matched C57BL6 mice. RESULTS: Deterioration in trabecular and cortical bone microarchitecture, reductions in bone formation, mineral apposition rate, bone volume/total volume, osteoid volume/bone surface, and bone strength were observed in Winnie mice compared with controls. Decreased osteoblast and increased osteoclast numbers were prominent in Winnie mice compared with controls. Upregulation of 5-HTR1B gene and increased association of FOXO1 with ATF4 complex were identified as associated mechanisms concomitant to overt inflammation and high levels of gut-derived serotonin in 14-week and 24-week Winnie mice. CONCLUSIONS: Skeletal phenotype of the Winnie mouse model of spontaneous chronic colitis closely represents manifestations of IBD-associated osteoporosis/osteopenia. The onset and progression of intestinal inflammation are associated with increased gut-derived serotonin level, increased bone resorption, and decreased bone formation.

The race for a COVID-19 vaccine: where are we up to?

INTRODUCTION: A novel strain of coronavirus, SARS-CoV-2, has triggered a global pandemic of coronavirus disease (COVID-19) in late 2019. In January 2020, the WHO declared this pandemic a public health emergency. This pandemic has already caused over 5.3 million deaths from more than 272 million infections. The development of a successful vaccine is an urgent global priority to halt the spread of SARS-CoV-2 and prevent further fatalities. Researchers are fast-tracking this process, and there have already been significant developments in preclinical and clinical phases in a relatively short period of time. Some vaccines have been approved either for emergency use or mass application in recent months. AREAS COVERED: Herein, we provide a general understanding of the fast-tracked clinical trial procedures and highlight recent successes in preclinical and clinical trials to generate a clearer picture of the progress of COVID-19 vaccine development. EXPERT OPINION: A good number of vaccines have been rolled out within a short period a feat unprecedented in medical history. However, the emergence of new variants over time has appeared as a new threat, and the number of infections and casualties is still on the rise and this is going to be an ongoing battle.

Immune to addiction: how immunotherapies can be used to combat methamphetamine addiction.

Introduction: The concept of anti-methamphetamine (METH) immunotherapies is a few decades old. A substantial amount of information has been generated on the development of anti-METH immunotherapies, particularly in the preclinical stages of development of vaccines and monoclonal antibody (mAb) treatments. However, the concept of treating METH use addiction with anti-METH immunotherapies is not well understood by many researchers or general readers. A series of questions commonly arise regarding the concept: how does it work? What is the antigen used? How exactly does the vaccine prevent METH addiction?Areas covered: This paper reviews the published articles relating to the mechanisms of METH use disorders, strategies used in the development of anti-METH immunotherapies, and the mechanism of action of these treatments. It provides clear explanations to questions surrounding the basis of anti-METH immunotherapies and contextualizes their development. It also identifies areas for future investigation to speed their translation into clinical use.Expert opinion: While METH immunotherapies, including vaccines and mAbs, have progressed significantly in the last 30 years, there are newer approaches that should be evaluated to improve their translatability. Approaches including nanoparticle vaccines, virus-like particles, and other novel methods should be fully evaluated as means of generating anti-METH immunity.

COVID-19 Vaccines in the Pipeline, Are Antibodies Adequate?

The COVID-19 pandemic caused by the novel coronavirus strain SARS-CoV-2 has already led to catastrophic consequences in global physical and psychological health, as well as economic recession [...].

Vaccine development against methamphetamine drug addiction.

INTRODUCTION: There are currently no effective treatments for Methamphetamine (METH) addiction and psychotherapy remains the sole treatment option. The development of immunopharmacotherapies for the treatment of drug addiction, overdose, and relapse management appears to be promising alternative and a significant body of information has been generated using various vaccine development strategies. Herein, we present an update on the developments toward anti-METH vaccines and their study outcomes in preclinical and clinical studies. AREAS COVERED: The scope of this article is to present an update on METH vaccine development strategies such as active vaccination through hapten design and the passive immunization through monoclonal antibodies along with preclinical and clinical studies. The relevant literatures and clinical trial outcomes were searched in databases including Google, Google Scholar, PubMed, Science Direct, ClinicalTrials.gov, and www.anzctr.org.au using specific keywords. EXPERT OPINION: Significant improvements have been developed for immunopharmacotherapies for METH addiction over the last two decades. However, only one monoclonal antibody candidate has been evaluated in a phase I clinical trial. At this moment, it is essential to evaluate the safety and efficacy of potential candidates in clinical trials to validate the importance of this platform drug-vaccine conjugation in order to manage or overcome METH addiction.

Microneedle Systems for Vaccine Delivery: the story so far.

INTRODUCTION: Vaccine delivery via a microneedle (MN) system has been identified as a potential alternative to conventional vaccine delivery. MN can be self-administered, is pain-free and is capable of producing superior immunogenicity. Over the last few decades, significant research has been carried out in this area, and this review aims to provide a comprehensive picture on the progress of this delivery platform. AREAS COVERED: This review highlights the potential role of skin as a vaccine delivery route using a microneedle system, examines recent advancements in microneedle fabrication techniques, and provides an update on potential preclinical and clinical studies on vaccine delivery through microneedle systems against various infectious diseases. Articles for the review study were searched electronically in PubMed, Google, Google Scholar, and Science Direct using specific keywords to cover the scope of the article. The advanced search strategy was employed to identify the most relevant articles. EXPERT OPINION: A significant number of MN mediated vaccine candidates have shown promising results in preclinical and clinical trials. The recent emergence of cleanroom free, 3D or additive manufacturing of MN systems and stability, together with the dose-sparing capacity of the Nanopatch® system, have made this platform, commercially, highly lucrative.