Identification of Proteins Involved in Airway Remodeling in Human Lung Fibroblasts

Asthma is an inflammatory disease associated with chronic perturbation of homeostatic mechanisms, leading to alterations in the structure of the airway walls, termed airway remodeling (AR). Lung fibrosis is characterized by increased deposition of extracellular matrix (ECM) proteins, especially collagens, and enhanced proliferation and activation of fibroblasts, and, ultimately, distortion of normal lung architecture and loss of respiratory function. L-arginine is a key precursor of nitric oxide and proline, an amino acid enriched in collagen. We hypothesized that L-arginine metabolism is altered in AR, ultimately affecting collagen synthesis. In this study we have analyzed the expression of key regulatory enzymes in the arginine pathway (Figure 1). Arginase, which converts L-arginine into L-ornithine and urea, is a key enzyme of the urea cycle in the liver, but it is also expressed in cells and tissues that lack a complete urea cycle, including the lung. Arginase exists as two distinct isoenzymes, arginase I and II, which are encoded by different genes. Arginase I is a cytosolic enzyme and is the predominant isoform in the liver, where it is highly expressed. Even though low levels of arginase II have been detected in liver as well, this mitochondrial enzyme is mainly expressed in extrahepatic tissue. Arginase isoenzymes and the downstream enzymes ornithine decarboxylase (ODC) and ornithine aminotransferase (OAT) are key regulators of critical processes associated with asthma including AR, cell hyperplasia and collagen deposition and have been implicated in the hyperplastic and fibrotic changes of AR, respectively. One of the biological functions of arginase II may be regulation of the synthesis of nitric oxide (NO) by means of competition with NO synthase (NOS) for the common substrate, L-arginine (Figure 1). In addition, extrahepatic arginase may be involved in tissue repair processes through the synthesis of L-ornithine. Thus, L-ornithine is a precursor of polyamines and L-proline, which are involved in cell proliferation and collagen synthesis, respectively. We investigated expressions of arginase I, arginase II, ODC, and OAT proteins by Western blot and cell immunochemistry in human lung fibroblast (HLF) from normal (NHLF) and diseased (DHLF) asthmatic donors

Resveratrol downregulates KIF15 in prostate cancer cells

Kinesin family member 15 (KIF15) is best known as a stabilizer for bipolar spindle formation during cell division. By interacting with the actin cytoskeleton, KIF15 is also involved in cell proliferation, apoptosis, and differentiation, and therefore implicated in several types of malignancies including liver, pancreatic, and breast cancers. In castration-resistant prostate cancer cells, KIF15 protects AR/AR-V7 from degradation by binding the N-terminus of AR/AR-V7 and enhancing the recruitment of deubiquitinating enzyme ubiquitin-specific protease 14 (USP14). On the other hand, KIF15 is upregulated by AR transcriptionally. We have previously reported that resveratrol (RSV) downregulates ARV7 in prostate cancer cells by enhancing ubiquitination-mediated ARV7 degradation. To understand if KIF15 plays a role in RSV-mediated ARV7 degradation, we treated 22RV1, an ARV7-positive cell line, with RSV and estimated the levels of KIF15 by Western blot assays. We found that RSV downregulates KIF15 in a dose (20uM-100uM) and time (24 or 48h)-dependent manner. Intriguingly, we also noticed some morphological changes when the cells were treated with RSV. Next, we will test if RSV-mediated ARV7 downregulation is KIF15-dependent. If so, we will explore whether RSV and KIF15 inhibitor inhibit RSV-positive prostate cancer cell proliferation additively or synergistically. We will also treat both 22RV1 and WPMY-1 cells (a normal prostate cell line) with RSV to see if RSV’s effect on morphological change is cancer cell-specific

Effects of Aspirin on Expression of Proteins Implicated in Airway Remodeling in Human Lung Fibroblasts

Abstract and poster under embargo until May 31, 2018

Emerging Therapeutic Strategies for COVID-19 Patients

Over 100,000 cases of COVID-19 patients infected with the novel coronavirus SARS-COV-2 have been reported worldwide in approximately 2 months, resulting in over 3000 deaths. Potential therapeutic strategies, including remdesivir, chloroquine phosphate, abidol, lopinavir/ritonavir, plasma, antibody, vaccine and stem cells are discussed in this review. With the number of patients increasing daily, there is an urgent need for effective therapeutic intervention

Modulation of Arginase-2 mRNA Levels by ω-3 PUFAs and Aspirin in Asthmatic Human Lung Fibroblasts

Airway remodeling (AR) increases disease severity, and morbidity of asthmatic patients by contributing to irreversible airflow obstruction and progressive declines in lung function. Arginase isoenzymes and the downstream enzymes ornithine decarboxylase (ODC) and ornithine aminotransferase (OAT) have been implicated in the hyperplastic and fibrotic changes of AR, respectively. Omega-3 polyunsaturated fatty acids (ω-3 PUFAs) and resolvin metabolites have anti-AR effects, but whether they are mediated through the arginase pathway is unclear. Our study intended to determine the effects of the ω-3 PUFAs eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), resolvin D1 (RvD1), TH1 cytokines, acetylsalicylic acid (ASA), cAMP, and dexamethasone (DEX) on the expression of arginase isoenzymes arginase 1 (ARG1) and arginase 2 (ARG2), ODC, and OAT in human lung fibroblasts (HLF) from normal (NHLF) and diseased (DHLF) asthmatic donors using reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Our data showed that EPA and EPA+DHA downregulated ARG2 mRNA 2-fold in both types of HLF. DHA, RvD1, and DEX did not alter mRNA levels for any of the genes studied. EPA lowered the ARG2 protein levels in DHLF, but did not affect those levels in NHLF. ASA upregulated ARG2 mRNA 5-fold and 7-fold in NHLF and DHLF, respectively, TH1 cytokines downregulated ARG2, ODC, and OAT mRNA in DHLF 10-fold, 2-fold, and 2.5-fold, respectively, and cAMP downregulated ARG2 mRNA 2-fold in DHLF. These results are the first to show a direct effect of ω-3 PUFAs on ARG2 mRNA levels and provide further evidence for a role of ω-3 PUFAs in AR

Resveratrol\u27s Effect on the Androgen Receptor Pathway as a Target for Decreasing Growth and Invasion of Melanoma and Methods of Application

Introduction: Melanoma is predicted to be the fifth most common cause of cancer mortality in the U.S. in 2021. Although melanoma affects both sexes, current literature shows it is more prevalent and more deadly in men. Several potential reasons for this difference have been proposed, but most literature describes a gap in male knowledge concerning skin cancer and unwillingness to go to the dermatologist as an explanation. That said, the androgen receptor has been proven to play a role in melanoma growth and metastasis, and may play a part in the difference in incidence and mortality rates between sexes. Resveratrol, a naturally occurring compound found in grapes and berries with poor bioavailability, has been shown to decrease melanoma cell growth in both in vitro and in vivo studies. Its effects on the expression of several genes involved in the androgen receptor pathway in non-melanoma cancer are well documented. Although several mechanisms have been proposed in which resveratrol effects melanoma cell growth, we suspect the androgen receptor pathway may be another target that warrants further investigation. Objective: Our objective is to reiterate the potential for resveratrol as a treatment in melanoma and to propose that it changes melanoma behavior at least partially through its interaction in the androgen receptor pathway. We also will discuss pros and cons of the various ways resveratrol may be delivered as a treatment for melanoma. Methods: To support our hypothesis we reviewed current literature and performed scratch wound assay, qPCR, and Western Blotting. Results: Melanoma cells treated with resveratrol showed a decrease in the distance traveled during scratch wound assay. Additionally, androgen receptor pathway gene expression and protein level was altered in cells treated with resveratrol. Conclusions: The androgen receptor pathway plays a role in melanoma cell growth and metastasis. Resveratrol is a potential therapeutic that decreases melanoma growth at least partially through its effects on this pathway. Further melanoma in vivo studies that explore the various methods of treatment with resveratrol are needed to help the transition from bench to bedside

Exploring the Applicability of Cisplatin Uptake Inhibitors in the Reduction/Prevention of Ototoxicity

Introduction: Cisplatin is a chemotherapeutic agent used to treat many solid tumors, including ovarian, testicular, bladder, lung, and head and neck tumors. The main side effects associated with the drug include nephrotoxicity, peripheral neurotoxicity, and ototoxicity, with ototoxicity being the dose-limiting effect. Objective: The study\u27s overall goal is to determine if cimetidine, a cisplatin uptake inhibitor, has protective effects against cisplatin cytotoxicity on hearing cells with minimal reduction in cisplatin\u27s chemotherapeutic effects on cancer cells. To accomplish this, we want to first determine cisplatin\u27s dose- and time-dependent effects on HEI-OC1 cell viability. Method: House Ear Institute-Organ of Corti 1 (HEI-OC1) cells were grown in 6-well plates and treated 24 hours later with cisplatin concentrations of either 0, 10, 50, 100, 150, or 200uM. Cells were imaged using an inverted microscope at 48 hours. The number of dead and alive cells was counted using an automated cell counter at 72 hours. Results: Increasing levels of cisplatin concentrations resulted in decreasing numbers of HEI-OC1 cells that were alive and showed changes to cell morphology. Conclusion: Cisplatin exerts a negative effect on HEI-OC1 cell viability. We will utilize the above-identified condition to treat cancer and HEI-OC1 cells side by side with and without cimetidine

The Effects of Resveratrol on Prostate Cancer through Targeting the Tumor Microenvironment.

Prostate cancer is one of the most common cancers diagnosed in men in the United States and the second leading cause of cancer-related deaths worldwide. Since over 60% of prostate cancer cases occur in men over 65 years of age, and this population will increase steadily in the coming years, prostate cancer will be a major cancer-related burden in the foreseeable future. Accumulating data from more recent research suggest that the tumor microenvironment (TME) plays a previously unrecognized role in every stage of cancer development, including initiation, proliferation, and metastasis. Prostate cancer is not only diagnosed in the late stages of life, but also progresses relatively slowly. This makes prostate cancer an ideal model system for exploring the potential of natural products as cancer prevention and/or treatment reagents because they usually act relatively slowly compared to most synthetic drugs. Resveratrol (RSV) is a naturally occurring stilbenoid and possesses strong anti-cancer properties with few adverse effects. Accumulating data from both in vitro and in vivo experiments indicate that RSV can interfere with prostate cancer initiation and progression by targeting the TME. Therefore, this review is aimed to summarize the recent advancement in RSV-inhibited prostate cancer initiation, proliferation, and metastasis as well as the underlying molecular mechanisms, with particular emphasis on the effect of RSV on TME. This will not only better our understanding of prostate cancer TMEs, but also pave the way for the development of RSV as a potential reagent for prostate cancer prevention and/or therapy

Resveratrol Enhances Polyubiquitination-Mediated ARV-7 Degradation in Prostate Cancer Cells

Although androgen deprivation therapy (ADT) serves as the primary treatment option for localized or metastatic prostate cancer, most cases eventually develop into castration-resistant prostate cancer (CRPC). However, androgen receptor (AR) continues to be functional in CRPC through various mechanisms, including the development of AR splicing variants, especially ARV7. Since it lacks the ligand binding domain but retains the intact DNA binding domain, ARV7 is constitutively active, which makes ARV7-positive prostate cancer responsive to neither abiraterone nor enzalutamide. In this study, we explored the effect of resveratrol on ARV7 transcriptional activity and the potential for development of resveratrol as a treatment for ARV7-positive prostate cancer. First, we ectopically expressed ARV7 in PC3 cells, an AR-negative prostate cancer cell line, and demonstrated that resveratrol is capable of inhibiting ARV7 transcriptional activity by downregulating ARV7 protein levels. Of note, resveratrol does not affect the mRNA levels of ARV7 nor its nuclear translocation. Next, we demonstrated that resveratrol is capable of downregulating the levels of the endogenously expressed ARV7 as well as AR target gene mRNAs in 22RV1 prostate cancer cells. Mechanistically, resveratrol downregulates ARV7 by enhancing ARV7 polyubiquitination and subsequent proteasome-mediated degradation. These findings suggest that resveratrol could be a potential treatment for ARV7-positive CPRC

Metformin Reverses Prostate Cancer Resistance to Enzalutamide by Targeting TGF-β1/STAT3 Axis-Regulated EMT.

Although the newly developed second-generation anti-androgen drug enzalutamide can repress prostate cancer progression significantly, it only extends the survival of prostate cancer patients by 4-6 months mainly due to the occurrence of enzalutamide resistance. Most of the previous studies on AR antagonist resistance have been focused on AR signaling. Therefore, the non-AR pathways on enzalutamide resistance remain largely unknown. By using C4-2, CWR22Rv1 and LNCaP cell lines, as well as mice bearing CWR22Rv1 xenografts treated with either enzalutamide or metformin alone or in combination, we demonstrated that metformin is capable of reversing enzalutamide resistance and restores sensitivity of CWR22Rv1 xenografts to enzalutamide. We showed that metformin alleviated resistance to enzalutamide by inhibiting EMT. Furthermore, based on the effect of metformin on the activation of STAT3 and expression of TGF-β1, we propose that metformin exerts its effects by targeting the TGF-β1/STAT3 axis. These findings suggest that combination of metformin with enzalutamide could be a more efficacious therapeutic strategy for the treatment of castration-resistant prostate cancer