Reprogramming of tumor-associated macrophages by metabolites generated from tumor microenvironment.

The tumor microenvironment comprises both tumor and non-tumor stromal cells, including tumor-associated macrophages (TAMs), endothelial cells, and carcinoma-associated fibroblasts. TAMs, major components of non-tumor stromal cells, play a crucial role in creating an immunosuppressive environment by releasing cytokines, chemokines, growth factors, and immune checkpoint proteins that inhibit T cell activity. During tumors develop, cancer cells release various mediators, including chemokines and metabolites, that recruit monocytes to infiltrate tumor tissues and subsequently induce an M2-like phenotype and tumor-promoting properties. Metabolites are often overlooked as metabolic waste or detoxification products but may contribute to TAM polarization. Furthermore, macrophages display a high degree of plasticity among immune cells in the tumor microenvironment, enabling them to either inhibit or facilitate cancer progression. Therefore, TAM-targeting has emerged as a promising strategy in tumor immunotherapy. This review provides an overview of multiple representative metabolites involved in TAM phenotypes, focusing on their role in pro-tumoral polarization of M2.

PCSK9 stimulates Syk, PKCδ, and NF-κB, leading to atherosclerosis progression independently of LDL receptor.

Proprotein convertase subtilisin/kexin type-9 (PCSK9) binds to and degrades low-density lipoprotein (LDL) receptor, leading to increase of LDL cholesterol in blood. Its blockers have emerged as promising therapeutics for cardiovascular diseases. Here we show that PCSK9 itself directly induces inflammation and aggravates atherosclerosis independently of the LDL receptor. PCSK9 exacerbates atherosclerosis in LDL receptor knockout mice. Adenylyl cyclase-associated protein 1 (CAP1) is the main binding partner of PCSK9 and indispensable for the inflammatory action of PCSK9, including induction of cytokines, Toll like receptor 4, and scavenger receptors, enhancing the uptake of oxidized LDL. We find spleen tyrosine kinase (Syk) and protein kinase C delta (PKCδ) to be the key mediators of inflammation after PCSK9-CAP1 binding. In human peripheral blood mononuclear cells, serum PCSK9 levels are positively correlated with Syk, PKCδ, and p65 phosphorylation. The CAP1-fragment crystallizable region (CAP1-Fc) mitigates PCSK9-mediated inflammatory signal transduction more than the PCSK9 blocking antibody evolocumab does.

Scoparone attenuates PD-L1 expression in human breast cancer cells by MKP-3 upregulation.

Breast cancer is a frequently occurring malignant tumor that is one of the leading causes of cancer-related deaths in women worldwide. Monoclonal antibodies that block programed cell death 1 (PD-1)/programed cell death ligand 1 (PD-L1) - a typical immune checkpoint - are currently the recommended standard therapies for many advanced and metastatic tumors such as triple-negative breast cancer. However, some patients develop drug resistance, leading to unfavorable treatment outcomes. Therefore, other approaches are required for anticancer treatments, such as downregulation of PD-L1 expression and promotion of degradation of PD-L1. Scoparone (SCO) is a bioactive compound isolated from Artemisia capillaris that exhibits antitumor activity. However, the effect of SCO on PD-L1 expression in cancer has not been confirmed yet. This study aimed to evaluate the role of SCO in PD-L1 expression in breast cancer cells in vitro. Our results show that SCO downregulated PD-L1 expression in a dose-dependent manner, via AKT inhibition. Interestingly, SCO treatment did not alter PTEN expression, but increased the expression of mitogen-activated protein kinase phosphatase-3 (MKP-3). In addition, the SCO-induced decrease in PD-L1 expression was reversed by siRNA-mediated MKP-3 knockdown. Collectively, these findings suggest that SCO inhibited the expression of PD-L1 in breast cancer cells by upregulating MKP-3 expression. Therefore, SCO may serve as an innovative combinatorial agent for cancer immunotherapy.

Association between unpredictable work schedule and work-family conflict in Korea.

Background: As unpredictable work schedule (UWS) has increased worldwide, various studies have been conducted on the resulting health effects on workers. However, research on the effect of UWS on workers' well-being in Korea is still insufficient. This study aimed to investigate the relationship between UWS and work-family conflict (WFC) using 6th Korean Working Conditions Survey (KWCS). Methods: Both UWS and WFC were measured using self-reported questionnaires, using data from the 6th KWCS conducted between 2020 and 2021, including 31,859 participants. UWS was measured by questions regarding the frequency of changes in work schedules and limited advanced notice. WFC was measured by questions regarding work to family and family to work conflicts. Logistic regression analysis was conducted to investigate the association between UWS and WFC. Results: The prevalence of UWS was higher among men, those under 40 years old, service and sales workers and blue-collar workers, and those with higher salaries. Workplace size also influenced UWS prevalence, with smaller workplaces (less than 50 employees) showing a higher prevalence. The odds ratio (OR) for WFC was significantly higher in workers with UWS compared to workers without UWS after adjusting for gender, age, marital status, occupation, salary, education, weekly working hours, shift work, company size, and having a child under the age of 18 years, employment status (OR: 3.71; 95% confidence interval: 3.23-4.25). Conclusions: The analysis of nationwide data revealed that UWS interferes with workers' performance of family roles, which can lead to WFC. Our findings suggest that it is crucial to implement policies to address unfair work schedule management, promoting a healthier work-life balance and fostering a conducive environment for family responsibilities.

A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury.

Animal models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) because of the lack of access to reliable sources of diseased human aortic valves. Among the various animal models, AVS rabbit models are one of the most commonly used in large animal studies. However, traditional AVS rabbit models require a long-term period of dietary supplementation and genetic manipulation to induce significant stenosis in the aortic valve, limiting their use in experimental studies. To address these limitations, a new AVS rabbit model is proposed, in which stenosis is induced by a direct balloon injury to the aortic valve. The present protocol describes a successful technique for inducing AVS in New Zealand white (NZW) rabbits, with step-by-step procedures for the preparation, the surgical procedure, and the post-operative care. This simple and reproducible model offers a promising approach for studying the initiation and progression of AVS and provides a valuable tool for investigating the underlying pathological mechanisms of the disease.

Association between shift work and serum homocysteine level in female electronic manufacturing services workers.

Background: Shift work has been shown to increase the risk of cardiovascular disease (CVD) based on several evidences. The classic risk factors of CVD include age, hypertension, smoking, obesity and diabetes. Recently, the serum homocysteine level has been reported to be a valuable indicator of CVD risk. This study aimed to determine the variation in serum homocysteine level as a cardiovascular risk indicator among female workers according to shift work. Methods: The data of regular health examination of workers at an electronic manufacturing services company in Yeongnam region, South Korea in 2019 were examined in this study. The investigation was based on a cross-sectional study conducted on 697 female workers (199 day workers and 498 shift workers). The sociodemographic and biochemical characteristics were compared between day workers and shift workers. Through a logistic regression analysis, the odds ratio (OR) of the increased serum homocysteine level in relation to shift work was determined. Results: Compared to female day workers, female shift workers showed significantly higher level of serum homocysteine (8.85 ± 2.16 vs. 9.42 ± 2.04 µmol/mL; p = 0.001). The OR of day workers against shift workers was 1.81 (95% confidence interval [CI]: 1.25-2.63). With the adjustment of variables that may influence the level of serum homocysteine, the adjusted OR was 1.68 (95% CI: 1.09-2.60). Conclusions: The serum homocysteine level was significantly higher in shift workers than in day workers. It is thus likely to be a useful predictor of CVD in shift workers.

Healing Predictors of Conservative Treatment for Juvenile Osteochondritis Dissecans of the Talus.

OBJECTIVE: To investigate the healing response of juvenile osteochondritis dissecans (JOCD) of the talus after conservative treatment, identify healing predictors, and develop a predictive model for healing. DESIGN: Retrospective study. SETTING: Clinics at a tertiary-level pediatric medical center. PATIENTS: Fifty-five patients (55 ankles) who presented with JOCD. INTERVENTIONS: Patients were managed with cast immobilization followed by activity restriction. MAIN OUTCOME MEASURES: The primary outcome measure of progressive lesion reossification was determined from the latest radiograph, after at least 6 months of nonoperative treatment. Final clinical evaluation was performed by a questionnaire and complementary telephone interview. Multivariate logistic regression was used to determine the influence of age, sex, lesion size, classification, location, duration of symptoms, containment lesion, and the occurrence of cyst-like lesions on healing potential. RESULTS: After nonoperative treatment, 18 (33%) of 55 lesions had failed to progress toward healing. An older age (P = 0.034) and a completely detached but undisplaced (grade III) lesion (P < 0.001) at the time of diagnosis were predictive for the failure of conservative treatment. A multivariate logistic regression best predictor model that included age and grade resulted in the best predicted healing and yielded an area under the curve of 0.920 (P < 0.001). CONCLUSION: In two-thirds of skeletally immature patients, conservative treatment resulted in the progressive healing of JOCD of the talus. For older patients with grade III lesions showing a lower healing probability, surgical treatment should be considered.

Association between coronavirus disease 2019-related workplace interventions and prevalence of depression and anxiety.

Background: Although coronavirus disease 2019 is causing a variety of psychological problems for workers, there are few longitudinal studies on changes in workers' mental health by workplace intervention. This study aimed to evaluate the change in the prevalence of depression and anxiety according to the active involvement of the workplace. Methods: This study was conducted on 1,978 workers at a workplace who underwent a health screening from January 2019 to August 2020, and classified depression and anxiety disorders using a self-report questionnaire. After the first pandemic, the company stopped health screening, took paid leave and telecommuting, and conducted interventions such as operating its own screening clinic. To see if this workplace intervention affects workers' mental health, we conducted generalized estimating equations to compare odds ratio (OR). Results: In the pre-intervention group, 384 people (16.86%) had depression, and 507 people (22.26%) had anxiety disorder. Based on the OR before intervention, the OR of depression decreased to 0.76 (0.66-0.87) and the OR of anxiety disorder decreased to 0.73 (0.65-0.82). Conclusions: As a result of this study, it was confirmed that workplace intervention was related to a decrease in depression and anxiety. This study provides basic data to improve workers' mental health according to workplace intervention, and further research is needed according to workplace intervention in the future.

Spatiotemporal dynamics of macrophage heterogeneity and a potential function of Trem2hi macrophages in infarcted hearts.

Heart failure (HF) is a frequent consequence of myocardial infarction (MI). Identification of the precise, time-dependent composition of inflammatory cells may provide clues for the establishment of new biomarkers and therapeutic approaches targeting post-MI HF. Here, we investigate the spatiotemporal dynamics of MI-associated immune cells in a mouse model of MI using spatial transcriptomics and single-cell RNA-sequencing (scRNA-seq). We identify twelve major immune cell populations; their proportions dynamically change after MI. Macrophages are the most abundant population at all-time points (>60%), except for day 1 post-MI. Trajectory inference analysis shows upregulation of Trem2 expression in macrophages during the late phase post-MI. In vivo injection of soluble Trem2 leads to significant functional and structural improvements in infarcted hearts. Our data contribute to a better understanding of MI-driven immune responses and further investigation to determine the regulatory factors of the Trem2 signaling pathway will aid the development of novel therapeutic strategies for post-MI HF.

Near-Infrared Light-Triggered Generation of Reactive Oxygen Species and Induction of Local Hyperthermia from Indocyanine Green Encapsulated Mesoporous Silica-Coated Graphene Oxide for Colorectal Cancer Therapy.

Near-infrared (NIR) light-mediated photothermal therapy (PTT) and photodynamic therapy (PDT) have widely been used for cancer treatment applications. However, a number of limitations (e.g., low NIR absorption capacity of photothermal agents, insufficient loading efficiency of photosensitive molecules) have hindered the widespread use of NIR-mediated cancer therapy. Therefore, we developed a mesoporous silica-coated reduced graphene oxide (rGO) nanocomposite that could provide a high encapsulation rate of indocyanine green (ICG) and enhance PTT/PDT efficiency in vitro and in vivo. The ICG-encapsulated nanocomposite not only enhances the photothermal effect but also generates a large number of tumor toxic reactive oxygen species (ROS). By conjugation of polyethylene glycol (PEG) with folic acid (FA) as a tumor targeting moiety, we confirmed that ICG-encapsulated mesoporous silica (MS)-coated rGO nanocomposite (ICG@MS-rGO-FA) exhibited high colloidal stability and intracellular uptake in folate receptor-expressing CT-26 colorectal cancer cells. Upon NIR laser irradiation, this ICG@MS-rGO-FA nanocomposite induced the apoptosis of only CT-26 cells via enhanced PTT and PDT effects without any damage to normal cells. Furthermore, the ICG@MS-rGO-FA nanocomposite revealed satisfactory tumor targeting and biocompatibility in CT-26 tumor-bearing mice, thereby enhancing the therapeutic effects of PTT and PDT in vivo. Therefore, this tumor-targeted ICG@MS-rGO-FA nanocomposite shows a great potential for phototherapy applications.

New Bone Formation in the Whole Decellularized Cortical Bone Scaffold Using the Model of Revitalizing a Haversian System.

ABSTRACT: Decellularized allogeneic bone chips act as scaffolds for bone tissue regeneration. Owing to their lack of osteogenic potentials compared to autologous bone graft, decellularized bone scaffolds (DBSs) have applied only to small partial bone defects in clinical settings. Furthermore, only decellularized cancellous bone chips have been limitedly used for the purpose of bone regeneration. The cortical bone has less porosity and less osteogenic materials such as bone morphogenetic proteins in comparison with cancellous bone. In this study, we tried to accelerate new bone formation within the decellularized cortical bone scaffold using a vascular pedicle as an in vivo bioreactor.Forty DBSs were divided into 4 groups with different conditionings (DBS+ demineralized bone matrix [DBM], DBS+DBM+me+mesenchymal stem cells, DBS+DBM+vascular pedicle, and DBS+DBM+vascular pedicle+mesenchymal stem cells) and implanted into the back of 5 rabbits. Half of the DBSs were examined at 8 weeks and the other half at 16 weeks to determine vascularization level and osteogenesis within each group. New bone formation and bone-forming cells related to osteogenesis were observed via histological staining. Inclusion of the vascular pedicle resulted in larger areas of bone regeneration. With time, osteon structures became more prominent in groups containing the vascular pedicle.In summary, vascularized DBSs combined with a vascular pedicle have shown promising results for bone regeneration, thereby representing potential therapeutic alternatives for autologous bone grafts or bone tissue free transfer in large or segmental bone defects. In addition, demineralized whole cortical bone matrix along with vascular pedicle and various bone inductive materials, such as DBM and recombinant human bone morphogenetic protein-2, may be an additional new option of an ideal osteoinductive system.

Nipple Reconstruction Using the Semilunar Flap and Omega-shaped Acellular Dermal Matrix Strut.

BACKGROUND: Various operative methods exist for nipple reconstruction. Selection of an appropriate skin flap and core strut material is imperative in achieving a satisfactory outcome in nipple reconstruction. Long-term maintenance of nipple projection requires further investigation by surgeons. We propose a new technique that uses a semilunar flap and omega-shaped acellular dermal matrix (ADM). METHODS: Total 53 nipples were reconstructed by this method. An omega-shaped ADM strut was inserted into the barrel made by a semilunar flap. The footplates of omega-shaped ADM struts were spread out under the subcutaneous tissue of the donor site of the semilunar flap to support the dome of the omega strut. RESULTS: The mean maintenance rate of nipple projection was 95.12 ± 6.30% at 3 weeks, 80.60 ± 8.93% at 3 months, and 71.70 ± 8.67% at 6 months postoperatively when compared to the projection observed in the immediate postoperative period. Thirty-five patients (66.0%) showed a maintenance rate over 70% at 6 months post operation, with most patients (94.3%) demonstrating a maintenance rate greater than 60%. CONCLUSIONS: Our study with the omega-shaped ADM strut showed superior maintenance rates of projection when compared to other studies on that used AlloDerm® as a core strut for nipple reconstruction. Omega-shaped struts, when made with cross-linked thick ADM, supported the skin flap quite well. We propose that our method combining the semilunar flap with an omega-shaped ADM may be a good option for nipple reconstruction. LEVEL OF EVIDENCE IV: "This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors   www.springer.com/00266 ."

Prevention of Breast Implant Displacement Using the Acellular Dermal Matrix Garter Belt.

BACKGROUND: Since the issue of breast implant-associated anaplastic large cell lymphoma, smooth breast implants tend to be the more preferred option in implant-based breast reconstructions, compared to its use previously. The most unfavorable aspect of smooth implants is lateral and inferior displacements, which are more common in patients who undergo breast reconstruction compared to augmentation mammoplasty. Hence, we introduce a prevention method for implant displacement using an acellular dermal matrix garter belt. METHODS: This study is a retrospective review of patients who had undergone implant-based breast reconstruction between April 2019 and December 2020. Some patients who have highly possibility of implant displacement, had undergone the application of an ADM garter belt to prevent lateral or superior displacement. Implant displacement was assessed before and at least 6 months postoperatively. RESULTS: A total of 155 IBR cases were recorded. ADM garter belts were applied in 27 patients (17.4%) who had a high tendency of implant displacement for several reasons, which could be classified into two categories: wide breast pocket (56%) and tight inferomedial breast pocket (44%). The intraoperative average distance of lateral slipping on patients' reconstructed breasts from the chest wall midline in supine position was 3.02 ± 0.81 cm and corrected to1.54 ± 0.69 cm at least 6 months postoperative follow-up. CONCLUSIONS: We utilized an ADM strap as an internalized garter belt to minimize implant displacement. This ADM garter belt combined with capsuloplasty might be an effective way to prevent the displacement of smooth implants in the patients with a greater risk of implant displacement. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors   www.springer.com/00266.

Copper arsenite-complexed Fenton-like nanoparticles as oxidative stress-amplifying anticancer agents.

We report copper(II) arsenite (CuAS)-integrated polymer micelles (CuAS-PMs) as a new class of Fenton-like catalytic nanosystem that can display reactive oxygen species (ROS)-manipulating anticancer therapeutic activity. CuAS-PMs were fabricated through metal-catechol chelation-based formation of the CuAS complex on the core domain of poly (ethylene glycol)-b-poly(3,4-dihydroxy-L-phenylalanine) (PEG-PDOPA) copolymer micelles. CuAS-PMs maintained structural robustness under serum conditions. The insoluble state of the CuAS complex was effectively retained at physiological pH, whereas, at endosomal pH, the CuAS complex was ionized to release arsenite and cuprous Fenton catalysts (Cu+ ions). Upon endocytosis, CuAS-PMs simultaneously released hydrogen peroxide (H2O2)-generating arsenite and Fenton-like reaction-catalyzing Cu+ ions in cancer cells, which synergistically elevated the level of highly cytotoxic hydroxyl radicals (•OH), thereby preferentially killing cancer cells. Animal experiments demonstrated that CuAS-PMs could effectively suppress the growth of solid tumors without systemic in vivo toxicity. The design rationale of CuAS-PMs may provide a promising strategy to develop diverse oxidative stress-amplifying agents with great potential in cancer-specific therapy.

Distribution of Lung-RADS categories according to job type in a single shipyard workers.

BACKGROUND: Recently, lung cancer screenings based on age and smoking history using low-dose computed tomography (LDCT) have begun in Korea. This study aimed to evaluate the distribution of lung imaging reporting and data system (Lung-RADS) categories in shipyard workers exposed to lung carcinogens such as nickel, chromium, and welding fumes according to job type, to provide basic data regarding indications for LDCT in shipyard workers. METHODS: This study included 6,326 workers from a single shipyard, who underwent health examinations with LDCT between January 2010 and December 2018. Data on age, smoking status and history, medical history, and job type were investigated. The participants were categorized into high-exposure, low-exposure, and non-exposure job groups based on the estimated exposure level of nickel, chromium, and welding fumes according to job type. Cox proportional hazard regression analysis was used to determine the difference between exposure groups in Lung-RADS category ≥ 3 (3, 4A, and 4B). RESULTS: Out of all participants, 97 (1.5%) participants were classified into Lung-RADS category ≥ 3 and 7 (0.1%) participants were confirmed as lung cancer. The positive predictive value (ratio of diagnosed lung cancer cases to Lung-RADS category ≥ 3) was 7.2%. The hazard ratio (HR) of Lung-RADS category ≥ 3 was 1.451 (95% confidence interval [CI]: 0.911-2.309) in low-exposure and 1.692 (95% CI: 1.007-2.843) in high-exposure job group. Adjusting for age and pack-years, the HR was statistically significant only in the high-exposure job group (HR: 1.689; 95% CI: 1.004-2.841). CONCLUSIONS: Based on LDCT and Lung-RADS, among male shipyard workers, Lung-RADS category ≥ 3 were significantly higher in the high-exposure job group. Their HR tended to be > 1.0 and was statistically significant in the high-exposure job group. Additional studies should be conducted to establish more elaborate LDCT indications for occupational health examination.

Atorvastatin and blood flow regulate expression of distinctive sets of genes in mouse carotid artery endothelium.

Hypercholesterolemia is a well-known pro-atherogenic risk factor and statin is the most effective anti-atherogenic drug that lowers blood cholesterol levels. However, despite systemic hypercholesterolemia, atherosclerosis preferentially occurs in arterial regions exposed to disturbed blood flow (d-flow), while the stable flow (s-flow) regions are spared. Given their predominant effects on endothelial function and atherosclerosis, we tested whether (1) statin and flow regulate the same or independent sets of genes and (2) statin can rescue d-flow-regulated genes in mouse artery endothelial cells in vivo. To test the hypotheses, C57BL/6 J mice (8-week-old male, n=5 per group) were pre-treated with atorvastatin (10mg/kg/day, Orally) or vehicle for 5 days. Thereafter, partial carotid ligation (PCL) surgery to induce d-flow in the left carotid artery (LCA) was performed, and statin or vehicle treatment was continued. The contralateral right carotid artery (RCA) remained exposed to s-flow to be used as the control. Two days or 2 weeks post-PCL surgery, endothelial-enriched RNAs from the LCAs and RCAs were collected and subjected to microarray gene expression analysis. Statin treatment in the s-flow condition (RCA+statin versus RCA+vehicle) altered the expression of 667 genes at 2-day and 187 genes at 2-week timepoint, respectively (P<0.05, fold change (FC)≥±1.5). Interestingly, statin treatment in the d-flow condition (LCA+statin versus LCA+vehicle) affected a limited number of genes: 113 and 75 differentially expressed genes at 2-day and 2-week timepoint, respectively (P<0.05, FC≥±1.5). In contrast, d-flow in the vehicle groups (LCA+vehicle versus RCA+vehicle) differentially regulated 4061 genes at 2-day and 3169 genes at 2-week timepoint, respectively (P<0.05, FC≥±1.5). Moreover, statin treatment did not reduce the number of flow-sensitive genes (LCA+statin versus RCA+statin) compared to the vehicle groups: 1825 genes at 2-day and 3788 genes at 2-week, respectively, were differentially regulated (P<0.05, FC≥±1.5). These results revealed that both statin and d-flow regulate expression of hundreds or thousands of arterial endothelial genes, respectively, in vivo. Further, statin and d-flow regulate independent sets of endothelial genes. Importantly, statin treatment did not reverse d-flow-regulated genes except for a small number of genes. These results suggest that both statin and flow play important independent roles in atherosclerosis development and highlight the need to consider their therapeutic implications for both.

In vivo MRI detection of intraplaque macrophages with biocompatible silica-coated iron oxide nanoparticles in murine atherosclerosis.

Identification of a vulnerable atherosclerotic plaque before rupture is an unmet clinical need. Integrating nanomedicine with multimodal imaging has the potential to precisely detect biological processes in atherosclerosis. We synthesized silica-coated iron oxide nanoparticles (SIONs) coated with rhodamine B isothiocyanate and polyethylene glycol and investigated their feasibility in the detection of macrophages in inflamed atherosclerotic plaques of apolipoprotein E-deficient (ApoE-/-) mice via magnetic resonance (MR) and fluorescence reflectance (FR) imaging. In vitro cellular uptake of SIONs was assessed in macrophages using confocal laser scanning microscopy (CLSM). In vivo MR imaging was performed 24 h after SION injection via the tail vein in 26-week-old ApoE-/- mice fed a high-cholesterol diet (HCD). We also performed FR imaging of the extracted aortas from four different mice: two normal-diet-fed C57BL/6 mice injected with saline or 10 mg/kg SIONs and two HCD-fed ApoE-/- mice injected with 5 or 10 mg/kg SIONs. The harvested aortas were cryosectioned and stained with immunohistochemical staining. The CLSM images at 24 h after incubation showed efficient uptake of SIONs by macrophages, with no evidence of cytotoxicity. The in vivo and ex vivo MR and FR images demonstrated SION deposition in the atheroma. Upon immunohistochemical staining of the aorta, CLSM images revealed colocalization of macrophages and SIONs in the atherosclerotic plaque. These results demonstrate that polyethylene glycosylated SIONs could be a highly effective method to identify macrophage activity in atherosclerotic plaques as a multimodal imaging agent.

AIMP3 induces laminopathy and senescence of vascular smooth muscle cells by reducing lamin A expression and leads to vascular aging in vivo.

Aminoacyl-tRNA synthetase-interacting multifunctional protein 3 (AIMP3), a tumor suppressor, mediates a progeroid phenotype in mice by downregulating lamin A. We investigated whether AIMP3 induces laminopathy and senescence of human aortic smooth muscle cells (HASMCs) and is associated with vascular aging in mice and humans in line with decreased lamin A expression. Cellular senescence was evaluated after transfecting HASMCs with AIMP3. Molecular analyses of genes encoding AIMP3, lamin A, chemokine (C-C motif) ligand 2 (CCL2), and C-C chemokine receptor type 2 (CCR2) and histological comparisons of aortas were performed with mice at various ages (7 weeks, 5 months, 12 months, 24 months, and 32 months), AIMP3-transgenic mice, and human femoral arteries of cadavers. AIMP3-transfected HASMCs exhibited increased AIMP3 and senescence marker p16 protein expression and decreased lamin A protein expression in accordance with their disrupted nuclear morphology in histological analyses. AIMP3-transgenic mice displayed increased AIMP3 protein expression and decreased lamin A protein expression in aortas together with typical aging pathologies. Similar changes were observed in wild-type aging (24-month-old) mice but not in wild-type young (7-week-old) mice. In humans, AIMP3 and lamin A protein expression was higher and lower, respectively, in femoral arteries of elderly individuals than in those of their younger counterparts. This study found that AIMP3 overexpression in vitro decreased lamin A expression and induced nuclear laminopathy and cellular senescence. Similar findings were made in the vasculature of aging mice and elderly humans.

Nanoparticle-Mediated Blocking of Excessive Inflammation for Prevention of Heart Failure Following Myocardial Infarction.

Severe cardiac damage following myocardial infarction (MI) causes excessive inflammation, which sustains tissue damage and often induces adverse cardiac remodeling toward cardiac function impairment and heart failure. Timely resolution of post-MI inflammation may prevent cardiac remodeling and development of heart failure. Cell therapy approaches for MI are time-consuming and costly, and have shown marginal efficacy in clinical trials. Here, nanoparticles targeting the immune system to attenuate excessive inflammation in infarcted myocardium are presented. Liposomal nanoparticles loaded with MI antigens and rapamycin (L-Ag/R) enable effective induction of tolerogenic dendritic cells presenting the antigens and subsequent induction of antigen-specific regulatory T cells (Tregs). Impressively, intradermal injection of L-Ag/R into acute MI mice attenuates inflammation in the myocardium by inducing Tregs and an inflammatory-to-reparative macrophage polarization, inhibits adverse cardiac remodeling, and improves cardiac function. Nanoparticle-mediated blocking of excessive inflammation in infarcted myocardium may be an effective intervention to prevent the development of post-MI heart failure.

Activation of Aryl Hydrocarbon Receptor by ITE Improves Cardiac Function in Mice After Myocardial Infarction.

Background The immune and inflammatory responses play a considerable role in left ventricular remodeling after myocardial infarction (MI). Binding of AhR (aryl hydrocarbon receptor) to its ligands modulates immune and inflammatory responses; however, the effects of AhR in the context of MI are unknown. Therefore, we evaluated the potential association between AhR and MI by treating mice with a nontoxic endogenous AhR ligand, ITE (2-[1'H-indole-3'-carbonyl]-thiazole-4-carboxylic acid methyl ester). We hypothesized that activation of AhR by ITE in MI mice would boost regulatory T-cell differentiation, modulate macrophage activity, and facilitate infarct healing. Methods and Results Acute MI was induced in C57BL/6 mice by ligation of the left anterior descending coronary artery. Then, the mice were randomized to daily intraperitoneal injection of ITE (200 µg/mouse, n=19) or vehicle (n=16) to examine the therapeutic effects of ITE during the postinfarct healing process. Echocardiographic and histopathological analyses revealed that ITE-treated mice exhibited significantly improved systolic function (P<0.001) and reduced infarct size compared with control mice (P<0.001). In addition, we found that ITE increased regulatory T cells in the mediastinal lymph node, spleen, and infarcted myocardium, and shifted the M1/M2 macrophage balance toward the M2 phenotype in vivo, which plays vital roles in the induction and resolution of inflammation after acute MI. In vitro, ITE expanded the Foxp3+ (forkhead box protein P3-positive) regulatory T cells and tolerogenic dendritic cell populations. Conclusions Activation of AhR by a nontoxic endogenous ligand, ITE, improves cardiac function after MI. Post-MI mice treated with ITE have a significantly lower risk of developing advanced left ventricular systolic dysfunction than nontreated mice. Thus, the results imply that ITE has a potential as a stimulator of cardiac repair after MI to prevent heart failure.