Neutralization of excessive CCL28 improves wound healing in diabetic mice.

Introduction: Chronic, non-healing skin wounds such as diabetic foot ulcers (DFUs) are common in patients with type 2 diabetes mellitus (T2DM) and often result in limb amputation and even death. However, mechanisms by which T2DM and inflammation negatively impact skin wound healing remains poorly understood. Here we investigate a mechanism by which an excessive level of chemokine CCL28, through its receptor CCR10, impairs wound healing in patients and mice with T2DM. Methods & Results: Firstly, a higher level of CCL28 was observed in skin and plasma in both patients with T2DM, and in obesity-induced type 2 diabetic db/db mice. Compared with WT mice, adipose tissue from db/db mice released 50% more CCL28, as well as 2- to 3-fold more IL-1ß, IL-6, and TNF-α, and less VEGF, as determined by ELISA measurements. Secondly, overexpression of CCL28 with adenovirus (Adv-CCL28) caused elevation of proinflammatory cytokines as well as CCR10 expression and also reduced eNOS expression in the dorsal skin of WT mice as compared with control Adv. Thirdly, topical application of neutralizing anti-CCL28 Ab dose-dependently accelerated wound closure and eNOS expression, and decreased IL-6 level, with an optimal dose of 1 µg/wound. In addition, mRNA levels of eNOS and anti-inflammatory cytokine IL-4 were increased as shown by real-time RT-PCR. The interaction between eNOS and CCR10 was significantly reduced in diabetic mouse wounds following application of the optimal dose of anti-CCL28 Ab, and eNOS expression increased. Finally, enhanced VEGF production and increased subdermal vessel density as indicated by CD31 immunostaining were also observed with anti-CCL28 Ab. Discussion: Taken together, topical application of neutralizing anti-CCL28 Ab improved dorsal skin wound healing by reducing CCR10 activation and inflammation in part by preventing eNOS downregulation, increasing VEGF production, and restoring angiogenesis. These results indicate anti-CCL28 Ab has significant potential as a therapeutic strategy for treatment of chronic non-healing diabetic skin wounds such as DFUs.

Influencing factors for thyroid function among radiation workers in Wuhan City / 预防医学

Objective@# To investigate the thyroid functions and influencing factors among radiation workers in Wuhan City, so as to provide insights into occupational health monitoring among radiation workers.@*Methods @#Radiation workers receiving physical examinations in Wuhan Prevention and Treatment Center for Occupational Diseases from January to October 2022 were enrolled, and participants' gender, age, smoking, alcohol consumption, medical history, medication use, types of occupational radiation and work duration were collected. Triiodothyronine (TT3), thyroxine (TT4), free thyroxine (FT4), free triiodothyronine (FT3) and thyroid stimulating hormone (TSH) were measured using a magnetic microparticle-based chemiluminescence immunoassay. Personnel dose equivalent was monitored using thermoluminescent dosimetry, and annual cumulative radiation dose was estimated. Factors affecting thyroid function were identified using a multivariable linear regression model.@*Results@#Totally 978 radiation workers were recruited, with a median age of 32.00 (interquartile range, 10.00) years, and including 782 men (79.96%) and 196 women (20.04%). There were 246 smokers (25.15%), 257 workers with alcohol consumption (26.28%) and 489 with a history of radiation work (50.00%). The median annual cumulative radiation dose was 0.20 (interquartile range, 0.24) mSv. The percentage of abnormal thyroid function was 14.72%. Multivariable logistic regression analysis showed that women (OR=1.925, 95%CI: 1.061-3.490), history of radiation work (OR=2.810, 95%CI: 1.119-7.057) and involving in medical application (OR=1.915, 95%CI: 1.101-3.332) were associated with abnormal thyroid function.@*Conclusions@#The percentage of abnormal thyroid function was 14.72% among radiation workers in Wuhan City. History of exposure to ionizing radiation, types of occupational radiation and gender were main factors affecting thyroid function.

Association of noise exposure, plasma microRNAs with arterial stiffness among Chinese workers.

Long-term noise exposure is reported to damage cardiovascular system, but the relationship between occupational noise exposure and arterial stiffness (AS) and the underlying mechanism is still unclear. We aimed to investigate the association of occupational noise exposure with arterial stiffness (AS), and further explore the mediation roles of microRNAs (miRNAs). A total of 838 workers were recruited from two companies in Wuhan, Hubei, China. Cumulative occupational noise exposure (CNE) was assessed through noise level of job title and work years in occupational noise. The AS for the participants were evaluated using brachial-ankle pulse wave velocity (baPWV) measured by an oscillometric device. Each 1-unit increase in CNE levels was significantly associated with a 0.002 (95% confidence interval (CI) = 0.001-0.003) unit increase in ln-transformed values of baPWV. In the sex-specific analysis, the association was significant in males (ß = 0.002, 95%CI = 0.001-0.003). Meanwhile, the risk of bilateral hearing loss at high frequency was significantly higher in the high-exposed group than non-exposed group (OR = 1.895, 95%CI = 1.024-3.508), and participants with bilateral hearing loss at high frequency had a significantly higher level of ln-transformed baPWV (ß = 0.032, 95%CI = 0.003-0.061). Occupational noise exposure and AS were both negatively associated with plasma miR-92a-3p and miR-21-5p, and the two miRNAs mediated 15.0% and 16.8% of the association of occupational noise with AS (P < 0.05). Our findings suggest that occupational noise exposure is positively associated with AS, and plasma miR-92a-3p and miR-21-5p may partly mediate such association.

Inhibition of CCL28/CCR10-Mediated eNOS Downregulation Improves Skin Wound Healing in the Obesity-Induced Mouse Model of Type 2 Diabetes.

Chronic, nonhealing skin wounds, such as diabetic foot ulcers (DFUs), are common in patients with type 2 diabetes. Here, we investigated the role of chemokine (C-C motif) ligand 28 (CCL28) and its receptor C-C chemokine receptor type 10 (CCR10) in downregulation of endothelial nitric (NO) oxide synthase (eNOS) in association with delayed skin wound healing in the db/db mouse model of type 2 diabetes. We observed reduced eNOS expression and elevated CCL28/CCR10 levels in dorsal skin of db/db mice and subdermal leg biopsy specimens from human subjects with type 2 diabetes. Further interrogation revealed that overexpression of CCR10 reduced eNOS expression, NO bioavailability, and tube formation of human dermal microvascular endothelial cells (HDMVECs) in vitro, which was recapitulated in mouse dorsal skin. In addition, incubation of HDMVECs with CCL28 led to internalization of the CCR10/eNOS complex and colocalization with lysosome-associated membrane protein 1. Finally, topical application of myristoylated CCR10 binding domain 7 amino acid (Myr-CBD7) peptide prevented CCR10-eNOS interaction and subsequent eNOS downregulation, enhanced eNOS/NO levels, eNOS/VEGF-R2+ microvessel density, and blood perfusion, reduced inflammatory cytokine levels, and importantly, decreased wound healing time in db/db mice. Thus, endothelial cell CCR10 activation in genetically obese mice with type 2 diabetes promotes eNOS depletion and endothelial dysfunction, and targeted disruption of CCR10/eNOS interaction improves wound healing.

Evaluating the capability of municipal solid waste separation in China based on AHP-EWM and BP neural network.

With the increase in municipal solid waste (MSW), most cities face solid waste management issues. In this study, the analytic hierarchy process (AHP) and artificial neural network (ANN) models were improved to assess the MSW separation capability based on 18 selected indicators of solid waste separation in 15 cities in China. The entropy weight method (EWM) was used in AHP to optimize and determine the indicators and then evaluate their weights, which showed that the general public budget expenditure had the highest weight (0.5239). This implied that the MSW separation capability could be mainly influenced by government financial support. ANN based on scan optimization and machine learning methods were established (R2 = 0.9992) to predict the missing indicators. The mapping relationship between MSW separation indicators and capabilities was also significantly improved from R2 = 0.5317 to R2 = 0.9993, thereby increasing the prediction accuracy of MSW separation capabilities to 95.15%. Thus, this research provides a new avenue for MSW separation and establishes a combined model to predict the separation capability in practical applications.

Stress hormone biosynthesis-based genes and lifestyle moderated the association of noise exposure with blood pressure in a cohort of Chinese tobacco factory workers: A cross-sectional analysis.

When evaluating noise-related cardiovascular risk, noise is generally solely assessed as the major stressor. However, cardiovascular effect of other simultaneous exposure events, such as unhealthy lifestyle and genetic variation, is easily neglected. The aim of this study is to estimate the combined effect of noise and lifestyle on blood pressure alteration, particularly under different genetic background. This study included 536 workers from a tobacco factory in Wuhan, China, who were divided into high exposure group and low exposure group according to noise measurement in their working area. All participants took annual physical examination and questionnaire survey to provide information on individual systolic and diastolic blood pressure (SBP and DBP) and lifestyle (smoking, drinking and physical activity). Single nucleotide polymorphism at genes related to stress hormone production were determined. Moderated moderation models were constructed to investigate the interaction effect of noise exposure and lifestyle factors on blood pressure with regard to different genetic background. We identified an expected trend in association between noise exposure and SBP among active smokers (P = 0.086). The moderated moderation analysis showed significant three-way interaction effect (COMT rs4680 × smoking status × noise exposure levels) on SBP or DBP (both P < 0.05). For COMT rs4680 GA+AA genotype carriers, active smoking significantly moderated the association between noise exposure and SBP or DBP (both P < 0.05). The results indicated that for COMT rs4680 A allele carriers, tobacco and noise exposure contribute collectively to blood pressure alteration, supporting that stress hormone production may play a certain role in the smoke-and-noise-induced cardiovascular effect.

Stem cells from a malignant rat prostate cell line generate prostate cancers in vivo: a model for prostate cancer stem cell propagated tumor growth.

Cancer stem cells (CSCs) are resistant to conventional cancer therapies, permitting the repopulation of new tumor growth and driving disease progression. Models for testing prostate CSC-propagated tumor growth are presently limited yet necessary for therapeutic advancement. Utilizing the congenic nontumorigenic NRP152 and tumorigenic NRP154 rat prostate epithelial cell lines, the present study investigated the self-renewal, differentiation, and regenerative abilities of prostate stem/progenitor cells and developed a CSC-based PCa model. NRP154 cells expressed reduced levels of tumor suppressor caveolin-1 and increased p-Src as compared to NRP152 cells. Gene knockdown of caveolin-1 in NRP152 cells upregulated p-Src, implicating their role as potential oncogenic mediators in NRP154 cells. A FACS-based Hoechst exclusion assay revealed a side population of stem-like cells (0.1%) in both NRP152 and NRP154 cell lines. Using a 3D Matrigel culture system, stem cells from both cell lines established prostaspheres at a 0.1% efficiency through asymmetric self-renewal and rapid proliferation of daughter progenitor cells. Spheres derived from both cell lines contained CD117+ and CD133+ stem cell subpopulations and basal progenitor cell subpopulations (p63+ and CK5+) but were negative for luminal cell CK8 markers at day 7. While some NRP152 sphere cells were androgen receptor (AR) positive at this timepoint, NRP154 cells were AR- up to 30 days of 3D culture. The regenerative capacity of the stem/progenitor cells was demonstrated by in vivo tissue recombination with urogenital sinus mesenchyme (UGM) and renal grafting in nude mice. While stem/progenitor cells from NRP152 spheroids generated normal prostate structures, CSCs and progeny cells from NRP154 tumoroids generated tumor tissues that were characterized by immunohistochemistry. Atypical hyperplasia and prostatic intraepithelial neoplasia (PIN) lesions progressed to adenocarcinoma with kidney invasion over 4 months. This provides clear evidence that prostate CSCs can repopulate new tumor growth outside the prostate gland that rapidly progresses to poorly differentiated adenocarcinoma with invasive capabilities. The dual in vitro/in vivo CSC model system presented herein provides a novel platform for screening therapeutic agents that target prostate CSCs for effective combined treatment protocols for local and advanced disease stages.

Association of noise exposure with risk of metabolic syndrome: Evidence from 44,698 individuals.

AIMS: Previous studies have explored the association between noise exposure and risk of metabolic syndrome (MetS); however, the results remain inconclusive. METHODS: PubMed and Web of Science databases were searched through December 2020, multivariate-adjusted relative risks (RRs) were pooled by using random-effects models. Subgroup analysis was also conducted stratifying by gender, study location, study design, source of noise, study quality, adjusting for smoking, drinking, body mass index, physical activity and shift work. RESULTS: Five studies involving 44,698 participants and 5187 MetS cases were included. A summarized adjusted RR for the relationship between noise exposure and risk of MetS was 1.27 (95% CI, 1.02-1.60), and 1.11 (1.02-1.21) for blood pressure and 1.11 (1.06-1.17) for blood glucose. Subgroup analysis revealed that the pooled risk of MetS was statistically significant in all cohort studies (RR = 1.34, 95 %CI, 1.06-1.68), ambient/traffic noise (RR = 1.24, 95 %CI, 1.13-1.35) and occupational noise by removing one low quality study (RR = 2.21, 95 %CI, 1.41-3.44). CONCLUSIONS: Noise exposure is associated with an increased risk of MetS, and occupational noise exposure may result in a greater risk. Additional more prospective large-scale studies conducted in more countries or populations are needed to confirm the results, establish causality and elucidate the underlying mechanisms.

Association of occupational noise exposure, bilateral hearing loss with atherosclerotic cardiovascular disease risk in Chinese adults.

We aimed to explore the association of occupational noise exposure with atherosclerotic cardiovascular disease (ASCVD) risk in Chinese adults. We included 21,412 participants from the Dongfeng-tongji Cohort Study, occupational noise exposure was evaluated through workplace noise level and/or the job titles, hearing loss was defined as a pure-tone mean of 25 dB or higher at 0.5, 1, 2, and 4 kHz in any ear. Compared with participants without occupational noise exposure, the 10-year ASCVD risk was significantly higher for noise exposure duration ≥20 years (OR = 1.20, 95%CI = 1.05-1.32) after adjusting for potential confounders. In the subgroup analysis, the association was only statistically significant in males (OR = 1.86, 95%CI = 1.12-3.14) and participants aged equal to or over 60 years old (OR = 1.20, 95%CI = 1.05-1.33), but not in females (OR = 1.15, 95%CI = 0.71-1.92) and aged below 60 (OR = 1.51, 95%CI = 0.75-2.85). In the subsample analyses (N = 10,165), bilateral hearing loss was associated with a higher risk of 10-year ASCVD (OR = 1.72, 95%CI = 1.30-2.30), especially for participants who were males (OR = 2.40, 95%CI = 1.61-3.42) and aged equal to or over 60 (OR = 1.85, 95%CI = 1.40-2.44). The present study suggests that occupational noise exposure may be a potential risk factor for ASCVD, especially for males and older participants.

Phagocytosis of apoptotic endothelial cells reprograms macrophages in skin wounds.

In healing wounds, the regression of blood vessels during the resolution phase creates a significant number of apoptotic endothelial cells (ApoECs). Surprisingly few studies have investigated the fate of apoECs in wounds, or the consequence of their removal. The current study employed both in vitro and in vivo models to investigate if macrophages ingest apoECs and to determine if such phagocytosis alters macrophage phenotype. To examine the capability of macrophages to ingest apoECs in in vivo wounds, pHrodo green labeled apoECs were injected into skin wounds 6 days after injury. The results demonstrated that 2.2% of macrophages in the wounds had engulfed apoECs 24 hours after injection. Macrophages that had engulfed apoECs expressed the markers CD80 (100%), CD86 (93.8%), and CD163 (22.8%), while no expression of CD206 marker was observed. In in vitro studies, 76.1% and 81.1% of PMA differentiated THP-1 macrophages engulfed apoECs at 6 and 24 hours, respectively. mRNA expression levels of IL-1ß, iNOS, and TGF-ß1 decreased in THP-1 macrophages after exposure to apoECs, while the expression of IL-6 increased. THP-1 macrophages that were incubated with apoECs for 6hours expressed CD80 (30.2%), CD163 (62.9%), and CD206 (45.3%), while expression levels in untreated group were 0.5%, 45.0%, and 2.4%, respectively. Taken together, our studies showed that macrophages phagocytize dermal apoECs both in vitro and in vivo. The engulfment of apoECs leads to a unique macrophage phenotype, which has characteristics of both M1 and M2 macrophage phenotypes. These findings provide a new mechanism by which macrophage phenotypes can be modified during wound resolution.

Dermal fibroblast phagocytosis of apoptotic cells: A novel pathway for wound resolution.

The effective clearance of apoptotic cells is an essential step in the resolution of healing wounds. In particular, blood vessel regression during wound resolution produces a significant number of apoptotic endothelial cells (ApoEC) that must be cleared. In considering the fate of ApoEC and the presence of fibroblasts during wound resolution, we hypothesized that fibroblasts might serve as phagocytes involved in endothelial cell removal. The current study investigated whether dermal fibroblasts engulf ApoEC, whether this uptake alters the phenotype of dermal fibroblasts, and the biological molecules involved. In both in vitro and in vivo studies, following ApoEC engulfment, fibroblasts acquired a pro-healing phenotype (increased cell migration, contractility, α-smooth muscle actin expression, and collagen deposition). In addition, fibroblast uptake of ApoEC was shown to be mediated in part by the milk fat globule-EGF factor 8 protein/integrin αv ß5 pathway. Our study demonstrates a novel function of fibroblasts in the clearance of ApoEC and suggests that this capability has significant implications for tissue repair and fibrosis.

A meta-analysis of observational studies including dose-response relationship between long working hours and risk of obesity.

We aimed to conduct a meta-analysis with stick reference and uniform cut-off of obesity to evaluate the relationship between long working hours and risk of obesity, using a quantitative dose-response method. The PubMed and Web of Science databases were searched through February 26, 2021, odds ratios (ORs) were pooled by using random-effects models and restricted cubic spline analysis with four knots was used to explore the dose-response relationship of working time and risk of obesity. Ten observational studies with 20 independent reports involving 189,590 participants were included in the present analysis. The summarized adjusted OR for the relationship between long working hours and obesity risk was 1.13 (95% CI: 1.01 to 1.26), when compared with weekly working hours less than and equal to 40 h using the random-effects model. A J-shaped association between long working hours and risk of obesity was observed (P < 0.001 for nonlinearity) for the dose-response relationship. Exclusion of any single study did not alter the combined relative risk. Individuals involved in long working hours are more likely to be obesity. Further studies are needed to confirm the results, and optimized and proper job arrangement should be established for improving the health of workers.

Prevalence of hearing loss and influencing factors among workers in Wuhan, China.

We aimed to estimate the prevalence of hearing loss and influencing factors among workers in automobile manufacturing industry in Wuhan, China. We conducted cross-sectional analyses of 2017 through 2019 data from survey of the key occupational diseases on 17,176 workers in automobile manufacturing industry, Wuhan, China. Hearing loss was defined as a pure tone mean of 25 dB or higher in either ear at 0.5, 1, and 2 kHz for speech frequency and at 3, 4, and 6 kHz for high frequency. Among the 17,176 workers, more than a quarter of participants had high frequency hearing loss, and 6.41% had speech frequency hearing loss. The prevalence of hearing loss was higher among participants with diabetes mellitus and current smoking, temporary tinnitus, and sudden change in hearing. Compared with the controls, age (OR = 1.08, 95% CI = 1.08-1.09), male (OR = 1.40, 95% CI = 1.21-1.63), occupational noise exposure (OR = 1.19, 95% CI = 1.08-1.30), having temporary tinnitus (OR = 1.20, 95% CI = 1.08-1.33), and having sudden change in hearing (OR = 1.58, 95% CI = 1.20-2.08) were associated with higher prevalence of high frequency hearing loss; meanwhile, age (OR = 1.09, 95% CI = 1.08-1.09), male (OR = 1.38, 95% CI = 1.11-1.71), having family history of hearing loss (OR = 2.84, 95% CI = 1.35-5.97), and having sudden change in hearing (OR = 2.58, 95% CI = 1.80-3.70) were associated with higher prevalence of speech frequency hearing loss. No additive and multiplicative interaction was found between occupational noise and these factors for hearing loss. Hearing loss directly affects 25% of workers in automobile manufacturing industry in Wuhan. Measures should be implemented for the control of occupational noise and other factors simultaneously in the workplace.

Is job strain associated with a higher risk of type 2 diabetes mellitus? A systematic review and meta-analysis of prospective cohort studies.

OBJECTIVES: Epidemiological studies have explored the relationship between work-related stress and the risk of type 2 diabetes mellitus (T2DM), but it remains unclear on whether work-related stress could increase the risk of T2DM. We aimed to evaluate the association between job strain and the risk of T2DM. METHODS: We searched PubMed and Web of Science up to April 2019. Summary risk estimates were calculated by random-effect models. And the analysis was also conducted stratifying by gender, study location, smoking, drinking, body mass index, physical activity, family history of T2DM, education and T2DM ascertainment. Studies with binary job strain and quadrants based on the job strain model were analyzed separately. RESULTS: A total of nine studies with 210 939 participants free of T2DM were included in this analysis. High job strain (high job demands and low control) was associated with the overall risk of T2DM compared with no job strain (all other combinations) [relative risk (RR) 1.16, 95% confidence interval (CI) 1.03-1.31], and the association was more evident in women (RR 1.48, 95% CI 1.02-2.14). A statistically significant association was also observed when using high strain as a category (job strain quadrants) rather than binary variable (RR 1.62, 95% CI 1.04-2.55) in women but not men. CONCLUSIONS: Our study suggests that job strain is an important risk factor for T2DM, especially among women. Appropriate preventive interventions in populations with high job strain would contribute to a reduction in T2DM risk.

Association of occupational noise exposure, bilateral hearing loss with hypertension among Chinese workers.

OBJECTIVE: To evaluate the relationship of occupational noise, bilateral hearing loss with blood pressure and hypertension among a Chinese population. METHODS: We included 15 422 individuals from a cross-sectional survey of the key occupational diseases in 2017 in Wuhan, Hubei Province, China. Occupational noise exposure was evaluated through workplace noise level and/or the job titles. Hearing loss was defined as a pure-tone average of 25 dB or higher at speech frequency (0.5, 1, 2 kHz) or high frequency (3, 4, 6 kHz) in both ears. Hypertension was defined as blood pressure at least 140/90 mmHg or self-reported current use of antihypertensive medication. RESULTS: Compared with participants without occupational noise exposure, the prevalence of hypertension was significantly higher for noise exposure duration of 5 to less than 10 years [odds ratio (OR) = 1.13, 95% confidence interval (CI) = 1.04-1.27] and at least 10 years (OR = 1.17, 95% CI = 1.09-1.30). In the sex-specific analysis, the association was significantly pronounced in male (OR = 1.18, 95% CI = 1.06-1.32 for duration of 5 to <10 years; OR = 1.25, 95% CI = 1.12-1.38 for duration ≥10 years), but not in female (OR = 1.01, 95% CI = 0.80-1.11 for duration of 5 to <10 years; OR = 1.06, 95% CI = 0.90-1.20 for duration ≥10 years). In the subsample analyses, bilateral hearing loss was associated with a higher prevalence of hypertension, no matter for speech frequency hearing loss (OR = 1.12, 95% CI = 1.02-1.30 for mild; OR = 1.35, 95% CI = 1.20-1.50 for severe) or for high-frequency hearing loss (OR = 1.24, 95% CI = 1.03-1.50 for mild; OR = 2.40, 95% CI = 1.80-3.17 for severe). The sex-subgroup analysis of hearing loss with hypertension was similar as occupational noise and hypertension. CONCLUSION: Our study has suggested occupational noise exposure is a potential risk factor for hypertension.

Live-Cell FRET Imaging of Phosphorylation-Dependent Caveolin-1 Switch.

The detection of dynamic conformational changes in proteins in live cells is challenging. Live-cell FRET (Förster Resonance Energy Transfer) is an example of a noninvasive technique that can be used to achieve this goal at nanometer resolution. FRET-based assays are dependent on the presence of fluorescent probes, such as CFP- and YFP-conjugated protein pairs. Here, we describe an experimental protocol in which live-cell FRET was used to measure conformational changes in caveolin-1 (Cav-1) oligomers on the surface of plasmalemma vesicles, or caveolae.

CCL28-induced CCR10/eNOS interaction in angiogenesis and skin wound healing.

Chemokines and their receptors play important roles in vascular homeostasis, development, and angiogenesis. Little is known regarding the molecular signaling mechanisms activated by CCL28 chemokine via its primary receptor CCR10 in endothelial cells (ECs). Here, we test the hypothesis that CCL28/CCR10 signaling plays an important role in regulating skin wound angiogenesis through endothelial nitric oxide synthase (eNOS)-dependent Src, PI3K, and MAPK signaling. We observed nitric oxide (NO) production in human primary ECs stimulated with exogenous CCL28, which also induced direct binding of CCR10 and eNOS resulting in inhibition of eNOS activity. Knockdown of CCR10 with siRNA lead to reduced eNOS expression and tube formation suggesting the involvement of CCR10 in EC angiogenesis. Based on this interaction, we engineered a myristoylated 7 amino acid CCR10-binding domain (Myr-CBD7) peptide and showed that this can block eNOS interaction with CCR10, but not with calmodulin, resulting in upregulation of eNOS activity. Importantly, topical administration of Myr-CBD7 peptide on mouse dermal wounds not only blocked CCR10-eNOS interaction, but also enhanced expression of eNOS, CD31, and IL-4 with reduction of CCL28 and IL-6 levels associated with improved wound healing. These results point to a potential therapeutic strategy to upregulate NO bioavailability, enhance angiogenesis, and improve wound healing by disrupting CCL28-activated CCR10-eNOS interaction.

Depletion of Caveolin-1 in Type 2 Diabetes Model Induces Alzheimer's Disease Pathology Precursors.

Type 2 diabetes mellitus (T2DM) is a risk factor for the development of late-onset Alzheimer's disease (AD). However, the mechanism underlying the development of late-onset AD is largely unknown. Here we show that levels of the endothelial-enriched protein caveolin-1 (Cav-1) are reduced in the brains of T2DM patients compared with healthy aging, and inversely correlated with levels of ß-amyloid (Aß). Depletion of Cav-1 is recapitulated in the brains of db/db (Leprdb ) diabetic mice and corresponds with recognition memory deficits as well as the upregulation of amyloid precursor protein (APP), BACE-1, a trending increase in ß-amyloid Aß42/40 ratio and hyperphosphorylated tau (p-tau) species. Importantly, we show that restoration of Cav-1 levels in the brains of male db/db mice using adenovirus overexpressing Cav-1 (AAV-Cav-1) rescues learning and memory deficits and reduces pathology (i.e., APP, BACE-1 and p-tau levels). Knocking down Cav-1 using shRNA in HEK cells expressing the familial AD-linked APPswe mutant variant upregulates APP, APP carboxyl terminal fragments, and Aß levels. In turn, rescue of Cav-1 levels restores APP metabolism. Together, these results suggest that Cav-1 regulates APP metabolism, and that depletion of Cav-1 in T2DM promotes the amyloidogenic processing of APP and hyperphosphorylation of tau. This may suggest that depletion of Cav-1 in T2DM underlies, at least in part, the development of AD and imply that restoration of Cav-1 may be a therapeutic target for diabetic-associated sporadic AD.SIGNIFICANCE STATEMENT More than 95% of the Alzheimer's patients have the sporadic late-onset form (LOAD). The cause for late-onset Alzheimer's disease is unknown. Patients with Type 2 diabetes mellitus have considerably higher incidence of cognitive decline and AD compared with the general population, suggesting a common mechanism. Here we show that the expression of caveolin-1 (Cav-1) is reduced in the brain in Type 2 diabetes mellitus. In turn, reduced Cav-1 levels induce AD-associated neuropathology and learning and memory deficits. Restoration of Cav-1 levels rescues these deficits. This study unravels signals underlying LOAD and suggests that restoration of Cav-1 may be an effective therapeutic target.