Hydrodynamic gene transfer of alpha-galactosidase (GLA) in the GLA knockout mouse partially reverses biochemical deficits

In humans, deficiency of the GLA enzyme is an inheritable, X-linked recessive disorder (Fabry disease) that results in accumulation of glycosphingolipid globotriaosylceramide (Gb3) within blood vessels, various tissues, and organs leading to severe pain, renal failure, cardiovascular disease and resultant premature mortality. Enzyme replacement therapy (ERT) is the only approved pharmacological treatment and is prohibitively expensive. Gene therapy may be a viable alternative to ERT. This study investigates the use of hydrodynamic gene delivery (HGD) of the GLA gene to GLA knockout mice, the Fabry disease animal model. The objective of this study is to evaluate the safety, efficacy and therapeutic outcome of administering GLA-containing plasmids via HGD to GLA knockout mice. The human GLA gene cDNA sequence was successfully inserted into the pAAV-MCS vector at the EcoR1 and Xho1 cloning sites resulting in pAAV-GLA plasmids and into the pLIVE vector at BamH1 and Xho1 sites resulting in pLIVE-GLA. It was determined that an 8% (of body weight) injection volume significantly increased GLA activity when compared to 6% (p<0.05) and injection volumes greater than 8% resulted in mortality. DNA doses of 5 to 15 µg/ml produced higher serum levels of GLA in the GLA knockout mice when compared to wild-type (WT) mice. The DNA dose used in this study was 15 µg/ml with an injection volume of 8%. Results show that pAAV-GLA generated greater activity (7600 %) in serum when compared to pLIVE-GLA (2500 %). With the pAAV-GLA plasmid, serum activity was maintained for two days. However, for pLIVE-GLA, at 18 days post-injection, GLA activity in liver and heart tissues in GLA knockout mice was significantly (p<0.05) increased when compared to WT. These levels were sustained for up to 66 days after the treatment when 3 treatments were administered, each 2 weeks apart. These data demonstrate that HGD of pLIVE-GLA safely and effectively corrected GLA deficiency in GLA knockout mice. Future studies should address Gb3 accumulation relative to GLA expression using the same model

Peculiarities of the function of cardiomyocytes in patients with ischemic heart disease associated with diabetes mellitus type 2

The functional activity of myocardium in patients with type 2 diabetes mellitus(T2DM) and coronary heart disease (CAD) was studied. The study included patients with CAD with T2DM as representatives of the main group, the comparison group consisted of patients with univariant course of ischemic heart disease without violation of carbohydrate metabolism. Contractile activity of the myocardium was evaluated by the inotroic reaction of isolated trabeculae of patients according to the Protocol tests "Post-rest" and "Ekstrasistoliei test". As a result, it was found that rhythm-inotropic myocardial response in patients with comorbid pathology persists to a greater extent than in the one-dimensional flow of CAD

Sensorimotor Experience Influences Recovery of Forelimb Abilities but Not Tissue Loss after Focal Cortical Compression in Adult Rats

Sensorimotor activity has been shown to play a key role in functional outcome after extensive brain damage. This study was aimed at assessing the influence of sensorimotor experience through subject-environment interactions on the time course of both lesion and gliosis volumes as well as on the recovery of forelimb sensorimotor abilities following focal cortical injury. The lesion consisted of a cortical compression targeting the forepaw representational area within the primary somatosensory cortex of adult rats. After the cortical lesion, rats were randomly subjected to various postlesion conditions: unilateral C5–C6 dorsal root transection depriving the contralateral cortex from forepaw somatosensory inputs, standard housing or an enriched environment promoting sensorimotor experience and social interactions. Behavioral tests were used to assess forelimb placement during locomotion, forelimb-use asymmetry, and forepaw tactile sensitivity. For each group, the time course of tissue loss was described and the gliosis volume over the first postoperative month was evaluated using an unbiased stereological method. Consistent with previous studies, recovery of behavioral abilities was found to depend on post-injury experience. Indeed, increased sensorimotor activity initiated early in an enriched environment induced a rapid and more complete behavioral recovery compared with standard housing. In contrast, severe deprivation of peripheral sensory inputs led to a delayed and only partial sensorimotor recovery. The dorsal rhizotomy was found to increase the perilesional gliosis in comparison to standard or enriched environments. These findings provide further evidence that early sensory experience has a beneficial influence on the onset and time course of functional recovery after focal brain injury

CERE-120 Prevents Irradiation-Induced Hypofunction and Restores Immune Homeostasis in Porcine Salivary Glands

Salivary gland hypofunction causes significant morbidity and loss of quality of life for head and neck cancer patients treated with radiotherapy. Preventing hypofunction is an unmet therapeutic need. We used an adeno-associated virus serotype 2 (AAV2) vector expressing the human neurotrophic factor neurturin (CERE-120) to treat murine submandibular glands either pre- or post-irradiation (IR). Treatment with CERE-120 pre-IR, not post-IR, prevented hypofunction. RNA sequencing (RNA-seq) analysis showed reduced gene expression associated with fibrosis and the innate and humoral immune responses. We then used a minipig model with CERE-120 treatment pre-IR and also compared outcomes of the contralateral non-IR gland. Analysis of gene expression, morphology, and immunostaining showed reduced IR-related immune responses and improved secretory mechanisms. CERE-120 prevented IR-induced hypofunction and restored immune homeostasis, and there was a coordinated contralateral gland response to either damage or treatment. CERE-120 gene therapy is a potential treatment for head and neck cancer patients to influence communication among neuronal, immune, and epithelial cells to prevent IR-induced salivary hypofunction and restore immune homeostasis

Adenovirus Gene Transfer to Amelogenesis Imperfecta Ameloblast-Like Cells

To explore gene therapy strategies for amelogenesis imperfecta (AI), a human ameloblast-like cell population was established from third molars of an AI-affected patient. These cells were characterized by expression of cytokeratin 14, major enamel proteins and alkaline phosphatase staining. Suboptimal transduction of the ameloblast-like cells by an adenovirus type 5 (Ad5) vector was consistent with lower levels of the coxsackie-and-adenovirus receptor (CAR) on those cells relative to CAR-positive A549 cells. To overcome CAR -deficiency, we evaluated capsid-modified Ad5 vectors with various genetic capsid modifications including “pK7” and/or “RGD” motif-containing short peptides incorporated in the capsid protein fiber as well as fiber chimera with the Ad serotype 3 (Ad3) fiber “knob” domain. All fiber modifications provided an augmented transduction of AI-ameloblasts, revealed following vector dose normalization in A549 cells with a superior effect (up to 404-fold) of pK7/RGD double modification. This robust infectivity enhancement occurred through vector binding to both αvβ3/αvβ5 integrins and heparan sulfate proteoglycans (HSPGs) highly expressed by AI-ameloblasts as revealed by gene transfer blocking experiments. This work thus not only pioneers establishment of human AI ameloblast-like cell population as a model for in vitro studies but also reveals an optimal infectivity-enhancement strategy for a potential Ad5 vector-mediated gene therapy for AI

Cellular and molecular mechanisms of immunomodulation in the brain through environmental enrichment

Recent studies on environmental enrichment (EE) have shown cytokines, cellular immune components [e.g., T lymphocytes, natural killer (NK) cells], and glial cells in causal relationship to EE in bringing out changes to neurobiology and behavior. The purpose of this review is to evaluate these neuroimmune mechanisms associated with neurobiological and behavioral changes in response to different EE methods. We systematically reviewed common research databases. After applying all inclusion and exclusion criteria, 328 articles remained for this review. Physical exercise (PE), a form of EE, elicits anti-inflammatory and neuromodulatory effects through interaction with several immune pathways including interleukin (IL)-6 secretion from muscle fibers, reduced expression of Toll-like receptors on monocytes and macrophages, reduced secretion of adipokines, modulation of hippocampal T cells, priming of microglia, and upregulation of mitogen-activated protein kinase phosphatase-1 in central nervous system. In contrast, immunomodulatory roles of other enrichment methods are not studied extensively. Nonetheless, studies showing reduction in the expression of IL-1β and tumor necrosis factor-α in response to enrichment with novel objects and accessories suggest anti-inflammatory effects of novel environment. Likewise, social enrichment, though considered a necessity for healthy behavior, results in immunosuppression in socially defeated animals. This has been attributed to reduction in T lymphocytes, NK cells and IL-10 in subordinate animals. EE through sensory stimuli has been investigated to a lesser extent and the effect on immune factors has not been evaluated yet. Discovery of this multidimensional relationship between immune system, brain functioning, and EE has paved a way toward formulating environ-immuno therapies for treating psychiatric illnesses with minimal use of pharmacotherapy. While the immunomodulatory role of PE has been evaluated extensively, more research is required to investigate neuroimmune changes associated with other enrichment methods.Gaurav Singhal, Emily J. Jaehne, Frances Corrigan and Bernhard T. Baun

Functional recovery following injury to the brain: Therapeutic effects of environmental stimulation

Mechanisms underlying functional recovery from traumatic brain injury (TBI) were investigated. In part one, the pattern of baseline metabolism and functional activation was studied following moderate TBI to define relevant loci for interventions aimed at promotion of functional recovery. In part two, animals were subjected to severe TBI and then placed in either standard cage housing or an enriched environment. Animals recovering from TBI in an enriched environment showed better behavioral outcomes and smaller lesion volumes as compared to their standard-housed counterparts. At two months post-TBI, animals showed severe metabolic abnormalities and total lack of functional activation in the whisker barrel circuit. In part three, the expression profiles of mRNA for three proteins (alpha-CAMKII, MAP2, Synaptophysin) associated with brain plasticity were studied following TBI with and without environmental enrichment. Analysis showed no differences between enriched and standard groups with respect to mRTNA expression profiles following TBI. It was found that alpha-CAMKII mRNA expression was significantly more perturbed following TBI than MAP2 or Synaptophysin. Taken together, these results shed further light on the complex picture of TBI pathophysiology and the mechanisms the brain utilizes to promote functional recovery following injury

Fall Prevention Nature Walk: Addressing Falls Prevention Across the Lifespan

Georgia Department of Public Health Injury Prevention Program (IPP) engages in robust state-based data and surveillance, strengthening strategic collaborations and partnerships, conducting assessments, and monitoring the effectiveness of evidence-based community programs. IPP is motivated to reduce risk factors for falls through a lens of shared risk and protective factors. By leveraging partnerships and resources we have developed an evidence-based fall prevention program that plays an important role in promoting health and well-being across the lifespan. According to the Centers for Disease Control and Prevention, falls are the second leading cause of unintentional injury deaths worldwide. In the United States, 28% of adults aged 65 and older report falling each year, resulting in approximately 36 million falls and 8 million fall related injuries each year. In Georgia, approximately 30.2% of adults aged 65 and older report falling each year. Despite the impact falls have on our communities, IPP acknowledges that falls are not an inevitable part of aging. Our approach to alleviate the problem is to support community-based efforts through the implementation of a Fall Prevention Nature Walk. This innovative approach derives from the Let’s Move Libraries StoryWalk® campaign used to promote literacy, reading, health, exercise, and movement in library settings across the U.S. The Fall Prevention Nature Walk engages participants (at every stage of life), through informational boards that incorporate evidence-based national resources, self-assessments, checklists, and community resources connected to fall risk and fall prevention. Twenty-four informational boards constitute a full set, and topics include self-advocacy, brain health, and age-appropriate exercise. The boards will be installed along paved paths in nature settings to promote movement and positive behavior changes. After the poster presentation from Georgia Department of Public Health Injury Prevention Program, attendees/learners will be able to identify risk factors for falls, discuss prevention strategies, and engage with local resources

Therapeutic Effects of Environmental Enrichment on Cognitive Function and Tissue Integrity Following Severe Traumatic Brain Injury in Rats

Postinjury environmental enrichment (EE) has been shown to alter functional and anatomical outcomes in a number of injury paradigms, including traumatic brain injury (TBI). The question of whether EE alters functional outcome following TBI in a model which produces overt histopathological consequences has not been addressed. We investigated this question using the severe, parasagittal fluid percussion injury (FPI) model. Rats (n = 7 per group, enriched and standard for behavior; n = 15 per group for histology) underwent severe (2.2–2.6 atm) FPI, with sham-operated rats (n = 7 per group, enriched and standard for behavior; n = 6 enriched, n = 3 standard for histology) serving as controls. Animals were allowed to recover for 11 days either in standard single housing or together (injured and sham) in an enriched environment consisting of a 92 × 61 × 77-cm ferret cage filled with various stimulatory objects. Consistent with earlier reports, injured animals recovering in the enriched environment showed significantly (P < 0.05) shorter latencies to find the platform in a Morris Water Maze task versus injured/standard animals on day 12 post-TBI. However, both injured groups showed significant deficits versus sham groups (P < 0.05). There were no differences between the sham/enriched and sham/standard groups. No significant group differences in swim speed were observed. At 14 days post-TBI, enriched animals had approximately twofold smaller lesion areas in regions of the cerebral cortex posterior to the injury epicenter (−4.5, −5.8, −6.8 mm relative to bregma; P < 0.05) compared to injured/standard animals. In addition, overall lesion volume for the entire injured cortical hemisphere was significantly smaller in animals recovering in the enriched environment. These results indicate that noninvasive environmental stimulation is beneficial in attenuating cognitive deficits and preserving tissue integrity in a TBI model which causes cerebral contusion and cell death