Browsing by Author "Vishwanatha, Jamboor K."
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Item A new approach to mentoring for research careers: the National Research Mentoring Network(BioMed Central Ltd., 2017-12-04) Sorkness, Christine A.; Pfund, Christine; Ofili, Elizabeth O.; Okuyemi, Kolawole S.; Vishwanatha, Jamboor K.; Team, NRMN; Zavala, Maria Elena; Pesavento, Theresa; Fernandez, Mary; Tissera, Anthony; Deveci, Alp; Javier, Damaris; Short, Alexis; Cooper, Paige; Jones, Harlan P.; Manson, Spero M.; Buchwald, Dedra S.; Eide, Kristin; Gouldy, Andrea; Kelly, Erin; Langford, Nicole; McGee, Richard; Steer, Clifford J.; Unold, Thad; Weber-Main, Anne Marie; Baez, Adriana; Stiles, Jonathan; Pemu, Priscilla; Thompson, Winston; Gwathmey, Judith; Lawson, Kimberly; Johnson, Japera; Hall, Meldra; Paulsen, Douglas; Fouad, Mona; Smith, Ann; Luna, Rafael; Wilson, Donald; Adelsberger, Greg; Simenson, Drew; Cook, Abby; Feliu-Mojer, Monica; Harwood, Eileen; Jones, Amy; Branchaw, Janet; Thomas, Stephen; Butz, Amanda; Byars-Winston, Angela; House, Stephanie; McDaniels, Melissa; Quinn, Sandra; Rogers, Jenna; Spencer, Kim; Utzerath, Emily; Duplicate Of, Weber-Main; Womack, VeronicaBACKGROUND AND PURPOSE: Effective mentorship is critical to the success of early stage investigators, and has been linked to enhanced mentee productivity, self-efficacy, and career satisfaction. The mission of the National Research Mentoring Network (NRMN) is to provide all trainees across the biomedical, behavioral, clinical, and social sciences with evidence-based mentorship and professional development programming that emphasizes the benefits and challenges of diversity, inclusivity, and culture within mentoring relationships, and more broadly the research workforce. The purpose of this paper is to describe the structure and activities of NRMN. KEY HIGHLIGHTS: NRMN serves as a national training hub for mentors and mentees striving to improve their relationships by better aligning expectations, promoting professional development, maintaining effective communication, addressing equity and inclusion, assessing understanding, fostering independence, and cultivating ethical behavior. Training is offered in-person at institutions, regional training, or national meetings, as well as via synchronous and asynchronous platforms; the growing training demand is being met by a cadre of NRMN Master Facilitators. NRMN offers career stage-focused coaching models for grant writing, and other professional development programs. NRMN partners with diverse stakeholders from the NIH-sponsored Diversity Program Consortium (DPC), as well as organizations outside the DPC to work synergistically towards common diversity goals. NRMN offers a virtual portal to the Network and all NRMN program offerings for mentees and mentors across career development stages. NRMNet provides access to a wide array of mentoring experiences and resources including MyNRMN, Guided Virtual Mentorship Program, news, training calendar, videos, and workshops. National scale and sustainability are being addressed by NRMN "Coaches-in-Training" offerings for more senior researchers to implement coaching models across the nation. "Shark Tanks" provide intensive review and coaching for early career health disparities investigators, focusing on grant writing for graduate students, postdoctoral trainees, and junior faculty. IMPLICATIONS: Partners from diverse perspectives are building the national capacity and sparking the institutional changes necessary to truly diversify and transform the biomedical research workforce. NRMN works to leverage resources towards the goals of sustainability, scalability, and expanded reach.Item Annexin A2 mediated TNBC metastasis via small extracellular vesicles: A multi-faceted omics-based characterization(2024-03-21) Trivedi, Rucha; Ranade, Payal; Vishwanatha, Jamboor K.Purpose – Triple-negative breast cancer (TNBC), the most aggressive subtype of breast cancer poses a high risk of metastasis due to a lack of targeted therapy and reliance on conventional treatment approaches. Among the organs susceptible to metastasis, the lungs are most frequently affected sites, accounting for approximately 60% of cases. Extracellular vesicles (EV) play a crucial role in establishing a pre-metastatic niche (PMN) for tumor cell homing to secondary sites. Identifying the tumor-derived EV-specific cargo that may potentially alter the lung microenvironment could establish an understanding of the molecular targets contributing to lung PMN development and consequently improving the poor TNBC prognosis and diagnosis. TNBC exhibits elevated expression of Annexin A2 (AnxA2), a plasma and endosome membrane-associated protein. The abundant expression of AnxA2 in tumor tissues and EVs derived from TNBC patients is correlated with poor overall survival and poor distant metastasis-free survival. In previous studies, depletion of AnxA2 protein levels in TNBC-derived EVs resulted in an altered PMN establishment, leading to significantly lower metastatic lesions in the lungs and brain in in vivo system. We aim to evaluate the molecular effects mediated by AnxA2 in TNBC-derived EVs and their cargo that aid the PMN formation and identify underlying molecular targets creating a favorable microenvironment by interaction with lung stroma. Methods and Results – To assess the role of AnxA2 in TNBC lung metastasis, we utilized a lung selective metastatic TNBC cell line, MDA MB 4175 (LM2). We used the shRNA-mediated stable knockdown technique to downregulate AnxA2 protein levels in LM2 cells. We then performed expressional analysis as well as its cell surface functionality using a plasmin generation assay. Additionally, we conducted tumor cell functional assays such as proliferation and invasion assays. Decreased AnxA2 protein levels in LM2 cells significantly reduced its plasmin generation, proliferative ability, and invasive potential. Subsequently, we isolated cell derived small EVs using differential ultracentrifugation and characterized them following the MISEV 2018 guidelines. We confirmed the purity, yield, size, presence of EV-enriched proteins (ESCRT, Hsp, CD9, CD81), and absence of negative proteins (Calnexin, GM130). We then subjected the EVs to quantitative proteomic analysis and identified the differentially expressed proteins upon AnxA2 depletion. We further exposed the healthy lung stromal fibroblasts and the local less-aggressive TNBC cells with EVs to assess the differences in local and distant site uptake and cargo transfer mediated by AnxA2. Conclusion – This comprehensive approach will determine the novel EV-associated proteins that act as functional regulators in promoting TNBC metastasis to the lungs. [This work is supported by the NCI R01CA220273 awarded to Dr. Jamboor K. Vishwanatha]Item Bacteriomimetic Nanoparticles for Immunotherapy against Breast Cancer(2015-03) Kokate, Rutika; Thamake, Sanjay; Chaudhary, Pankaj; Raut, Sangram; Mott, Brittney; Vishwanatha, Jamboor K.Short description: Immunotherapy represents a potential and innovative means to combat cancer. It essentially harnesses the body’s immune system to fight against cancer. Previous literature suggests that cancer vaccines designed against a specific tumor antigen have been efficiently utilized to trigger immune responses against tumor cells. Despite the preliminary evidence in animal models, low immunogenicity is one of the major hurdles in the development of vaccines in humans. In order to surmount this obstacle, several approaches including the use of an “ideal” tumor antigen, appropriate delivery techniques, immune boosting strategies with co-stimulatory molecules are being explored. Purpose: The purpose of this study was to develop “bacteriomimetic nanoparticles” to enhance adaptive cell-mediated immune responses (CD4+ and CD8+ T cell responses) against tumor antigen as a therapeutic option for cancer treatment. Materials and Methods: NPs were prepared by modified solid/oil/water solvent evaporation method using an ultrasonic processor UP200H system (Hielscher Ultrasonics GmbH, Germany). We used membrane preparations of the 4T1 mouse mammary cancer cell line as a tumor antigen and CpG ODN’s as a “bacteriomimetic” stimulant. Fourteen days before tumor challenge BALB/c female mice (6-8 weeks) were pre-immunized with CpG followed by secondary immunization using respective NPs encapsulated with the membrane antigen preparation. Subsequently, mice (n=5) were challenged subcutaneously (SC) with 105 tumor cells and the primary tumor size was monitored using vernier caliper and bioluminiscence imaging (Caliper Life Sciences Inc., MA, USA). Mice were sacrificed on day fourteen after tumor challenge; spleen cells were used for flow cytometric analysis and primary tumor tissue was used to evaluate effect of NP immunization on tumor growth, survival as well as the immune response (CD4+ and CD8+ T cell) via immunohistochemistry. Results: We found significant reduction in progression of tumor growth in mice immunized with CpG coated NPs containing tumor antigen (CpG-NP-Tag). Histological analysis confirmed that tumors in CpG-NP-Tag mice were relatively well differentiated and of lower grade in contrast to CpG-Blank tumors. Immunofluorescence (IF) data further revealed that CpG-NP-Tag tumors had lesser proliferation and higher apoptotic activity. Tumor CD4+ and CD8+T cell infiltration as well as T cell response in spleen was found be higher in CpG-NP-Tag NP immunized mice as compared to the controls (CpG-NP-Blank and NP-Tag). Conclusions: Primary tumor size, IHC, IF and flow cytometry analysis indicate that CpG-NP-Tag NPs were successfully employed to boost the immune response against tumor cells.Item Bioengineered Nanoparticles for Targeted Cancer Therapy(2018-05) Gdowski, Andrew S.; Vishwanatha, Jamboor K.; Ranjan, Amalendu P.; Cistola, David P.; Olivencia-Yurvati, Albert H.Despite improved overall survival in cancer patients over the past 50 years, limited advances have been made in treating patients with metastatic cancers. Multiple types of cancers demonstrate the unique ability to specifically metastasize to the bone. Among these, prostate cancer exhibits increased capacity to create bone specific lesions with high frequency. Once bone localization takes place, treatment regimens are limited and overall survival is poor. These bone metastases often cause debilitating and life threatening problems including: uncontrollable pain, hypercalcemia, broken bones, spinal cord compression, and the inability to perform activities of daily living. The overarching goal of this thesis was to develop novel bone targeted nanoparticle therapies. The first generation of nanoparticles we engineered and tested were designed to target the hydroxyapatite structure of the bone particularly in areas of high bone turnover with subsequent therapeutic release at the site of the tumor. Notably, this nanoparticle formulation was efficacious in decreasing prostate cancer bone metastatic tumors, improving bone structure, and reducing pain in a mouse model. The next generation of nanoparticles were developed to simultaneously target the bone endothelium and tumor cells using a programmable bioinspired approach with guidance from genomic information of prostate cancer patients. This novel bioinspired nanoparticle demonstrated enhanced ability to self-recognize cancer cells as well as improved bone homing and retention in our in vivo evaluation. Finally, we addressed the challenge of nanoparticle manufacturing scale up from lab size quantities to large scale batches using a microfluidic process. It is our sincere hope that concepts and publications derived from this thesis will help guide future efforts for targeted therapy and improve the lives of patients with cancer.Item Cabazitaxel-Loaded Nanoparticles Reduce the Invasiveness in Metastatic Prostate Cancer Cells: Beyond the Classical Taxane Function(MDPI, 2023-02-26) Lampe, Jana B.; Desai, Priyanka P.; Tripathi, Amit K.; Sabnis, Nirupama A.; Chen, Zhe; Ranjan, Amalendu P.; Vishwanatha, Jamboor K.Bone-metastatic prostate cancer symbolizes the beginning of the later stages of the disease. We designed a cabazitaxel-loaded, poly (lactic-co-glycolic acid) (PLGA) nanoparticle using an emulsion-diffusion-evaporation technique. Bis (sulfosuccinimidyl) suberate (BS3) was non-covalently inserted into the nanoparticle as a linker for the conjugation of a bone-targeting moiety to the outside of the nanoparticle. We hypothesized that the nanoparticles would have the ability to inhibit the epithelial-to-mesenchymal transition (EMT), invasion, and migration in prostate cancer cells. Targeted, cabazitaxel-loaded nanoparticles attenuated the EMT marker, Vimentin, and led to an increased E-cadherin expression. These changes impart epithelial characteristics and inhibit invasive properties in cancer progression. Consequently, progression to distant sites is also mitigated. We observed the reduction of phosphorylated Src at tyrosine 416, along with increased expression of phosphorylated cofilin at serine 3. These changes could affect migration and invasion pathways in cancer cells. Both increased p-120 catenin and inhibition in IL-8 expression were seen in targeted, cabazitaxel-loaded nanoparticles. Overall, our data show that the targeted, cabazitaxel-loaded nanoparticles can act as a promising treatment for metastatic prostate cancer by inhibiting EMT, invasion, and migration, in prostate cancer cells.Item CELL SURFACE TRANSLOCATION OF ANNEXIN A2 FACILITATES GLUTAMATE-INDUCED EXTRACELLULAR PROTEOLYSIS(2014-03) Maji, Sayantan; Vishwanatha, Jamboor K.; Valapala, MallikaNeurodegenerative diseases like age related macular degeneration (AMD) and Retinitis Pigmentosa (RP) are major causes of blindness affecting millions of people around the world. One of the major reasons of cell death observed in these diseases is the increased accumulation of glutamate, an excitatory amino acid. Unfortunately, the mechanisms behind glutamate induced toxicity are not yet known. Here we are investigating a possible role of a protein Annexin A2 (AnxA2) in glutamate induced toxicity. We found that glutamate causes increased membrane translocation of AnxA2. Increased membrane localization of AnxA2 and thereby its function can lead to the death of the eye cells leading to degenerative diseases like AMD and RP. The present study shows one of the possible mechanisms that can lead to glutamate induced cell death of the eye. Thus, using AnxA2 targeted therapy as an adjunctive therapy can lead to better and more efficient outcomes. Purpose (a): Glutamate-induced intracellular increase in Ca2+ levels leads to the hyper-activation of several normal Ca2+-mediated physiological processes including the activation of intracellular kinases, phosphatases, phospholipases and proteases which contribute to the degeneration of the retinal neurons as seen in many diseases including age-related macular degeneration (AMD) and retinitis pigmentosa (RP). Despite intensive research, the mechanisms that contribute to glutamate-induced cellular loss are yet to be elucidated. AnxA2, a Ca2+-dependent phospholipid binding protein serves as an extracellular proteolytic center by recruiting tissue plasminogen activator and plasminogen, and mediating localized generation of plasmin. We investigated whether AnxA2 plays a major role in glutamate induced neuronal excitotoxicity in a cone-photoreceptor cell line, 661W. Understanding the molecular mechanisms of glutamate-induced retinal degeneration can lead to the development of better therapeutic approaches for neurodegenerative diseases including AMD and RP. Our study provides new insights into one of the mechanisms that might contribute to glutamate-induced loss of photoreceptors in the retina. Methods (b): Ratiometric Ca2+ imaging and time lapse confocal microscopy were used to study glutamate-induced Ca2+ influx. EDTA eluates of 661W cells were immunoblotted to study the membrane translocation of endogenous as well as AnxA2-GFP in the presence or absence of different treatments. To determine whether glutamate induced membrane translocation of AnxA2 is dependent on the phosphorylation of the 23rd tyrosine residue or not, phosphomimetic and non-phosphomimetic variants were studied. Results (c): Glutamate translocated both endogenous and AnxA2-GFP to the cell surface in a process dependent on the activity of the NMDA receptor. Glutamate-induced translocation of AnxA2 is dependent on the phosphorylation of tyrosine 23 at the N-terminus and mutation of tyrosine 23 to a non-phosphomimetic variant inhibits the translocation process. The cell surface translocated AnxA2 forms an active plasmin-generating complex and this activity can be neutralized by a hexapeptide directed against the N-terminus. Conclusions (d): These results suggest an involvement of AnxA2 in potentiating glutamate-induced cell death processes. Thereby, targeting AnxA2 can be used as an adjunctive therapy in neurodegenerative diseases like AMD and RP.Item Clinical Significance of Annexin A2 in Predicting Poor Prognosis in African American Women with Triple-Negative Breast Cancer(2017-05) Gibbs, Lee D.; Vishwanatha, Jamboor K.; Basha, Riyaz; Lovely, Rehana S.; Mathew, Porunelloor A.; Singh, MeharvanTriple-negative breast cancers (TNBC) are identified by the absence of these three major receptors that drive most breast cancer subtypes. TNBC is the most aggressive breast cancer subtype and studies have shown that the incidence of TNBC is much higher in premenopausal African American (AA) women and woman of African descent in comparison to woman of European descent. TNBC in AA women has been associated with worst overall survival after controlling for socioeconomic factors, treatment latency, and tumor receptor expression. This suggests that the clinical outcome of TNBC in AA women may result more from biological differences than access to adequate healthcare. Utilizing a large archived breast cancer cohort of genome sequencing information and the evaluation of these targets in breast tissue and serum can lead to recognition of reliable biological markers that have tremendous potential to enhance detection, treatment, and prognosis. Our previous studies have shown that Annexin A2 (AnxA2), a 36 kda calcium-dependent phospholipid binding protein, is abundantly expressed in TNBC. We have shown AnxA2 to play multiple roles in TNBC by regulating cellular functions; including plasminogen activation, angiogenesis, proliferation, migration, invasion, and metastasis. AnxA2 is one of the most identified proteins expressed in exosomes (small vesicles that are secreted from tumors as metastatic regulators). We have previously demonstrated exosomal AnxA2 contribution to metastasis of TNBC cells in vivo. The proposed study will determine the correlation of AnxA2 with poor prognoses in AA TNBC patients, and establish the clinical significance of exosomal AnxA2 in contributing to the poor clinical outcomes seen in AA TNBC patients. Three specific aims were addressed in this work. Aim 1- Determine the association of secreted exosomal AnxA2 with TNBC amongst AA patients. Aim 2 - Evaluate AnxA2 expression in TNBC tissue samples amongst a breast cancer patient cohort of various breast subtypes. Aim 3 - Determine the correlation of AnxA2 gene expression with poor pathological, prognostic variables and race/ethnicity in TNBC patients through in silico analysis.Item Combination of Small Extracellular Vesicle-Derived Annexin A2 Protein and mRNA as a Potential Predictive Biomarker for Chemotherapy Responsiveness in Aggressive Triple-Negative Breast Cancer(MDPI, 2023-01-09) Desai, Priyanka P.; Narra, Kalyani; James, Johanna D.; Jones, Harlan P.; Tripathi, Amit K.; Vishwanatha, Jamboor K.Small extracellular vesicles (sEVs), mainly exosomes, are nanovesicles that shed from the membrane as intraluminal vesicles of the multivesicular bodies, serve as vehicles that carry cargo influential in modulating the tumor microenvironment for the multi-step process of cancer metastasis. Annexin A2 (AnxA2), a calcium(Ca(2+))-dependent phospholipid-binding protein, is among sEV cargoes. sEV-derived AnxA2 (sEV-AnxA2) protein is involved in the process of metastasis in triple-negative breast cancer (TNBC). The objective of the current study is to determine whether sEV-AnxA2 protein and/or mRNA could be a useful biomarkers to predict the responsiveness of chemotherapy in TNBC. Removal of Immunoglobulin G (IgG) from the serum as well as using the System Bioscience's ExoQuick Ultra kit resulted in efficient sEV isolation and detection of sEV-AnxA2 protein and mRNA compared to the ultracentrifugation method. The standardized method was applied to the twenty TNBC patient sera for sEV isolation. High levels of sEV-AnxA2 protein and/or mRNA were associated with stage 3 and above in TNBC. Four patients who responded to neoadjuvant chemotherapy had high expression of AnxA2 protein and/or mRNA in sEVs, while other four who did not respond to chemotherapy had low levels of AnxA2 protein and mRNA in sEVs. Our data suggest that the sEV-AnxA2 protein and mRNA could be a combined predictive biomarker for responsiveness to chemotherapy in aggressive TNBC.Item Community perspectives on AI/ML and health equity: AIM-AHEAD nationwide stakeholder listening sessions(PLOS, 2023-06-30) Vishwanatha, Jamboor K.; Christian, Allison; Sambamoorthi, Usha; Thompson, Erika L.; Stinson, Katie; Syed, Toufeeq A.Artificial intelligence and machine learning (AI/ML) tools have the potential to improve health equity. However, many historically underrepresented communities have not been engaged in AI/ML training, research, and infrastructure development. Therefore, AIM-AHEAD (Artificial Intelligence/Machine Learning Consortium to Advance Health Equity and Researcher Diversity) seeks to increase participation and engagement of researchers and communities through mutually beneficial partnerships. The purpose of this paper is to summarize feedback from listening sessions conducted by the AIM-AHEAD Coordinating Center in February 2022, titled the "AIM-AHEAD Community Building Convention (ACBC)." A total of six listening sessions were held over three days. A total of 977 people registered with AIM-AHEAD to attend ACBC and 557 individuals attended the listening sessions across stakeholder groups. Facilitators led the conversation based on a series of guiding questions, and responses were captured through voice and chat via the Slido platform. A professional third-party provider transcribed the audio. Qualitative analysis included data from transcripts and chat logs. Thematic analysis was then used to identify common and unique themes across all transcripts. Six main themes arose from the sessions. Attendees felt that storytelling would be a powerful tool in communicating the impact of AI/ML in promoting health equity, trust building is vital and can be fostered through existing trusted relationships, and diverse communities should be involved every step of the way. Attendees shared a wealth of information that will guide AIM-AHEAD's future activities. The sessions highlighted the need for researchers to translate AI/ML concepts into vignettes that are digestible to the larger public, the importance of diversity, and how open-science platforms can be used to encourage multi-disciplinary collaboration. While the sessions confirmed some of the existing barriers in applying AI/ML for health equity, they also offered new insights that were captured in the six themes.Item Correction to: Serum exosomal-annexin A2 is associated with African-American triple-negative breast cancer and promotes angiogenesis(BioMed Central Ltd., 2020-03-23) Chaudhary, Pankaj; Gibbs, Lee D.; Maji, Sayantan; Lewis, Cheryl M.; Suzuki, Sumihiro; Vishwanatha, Jamboor K.After publication of the original article [1], we were notified that the wrong version of Fig. 2b has been published.Item COVID-19 clinical trial participation and awareness in Texas(Informa UK Limited, trading as Taylor & Francis Group, 2024-04-25) Luningham, Justin M.; Akpan, Idara N.; Alkhatib, Sarah; Taskin, Tanjila; Desai, Palak; Vishwanatha, Jamboor K.; Thompson, Erika L.The COVID-19 pandemic required the rapid development of COVID-19 vaccines and treatments, necessitating quick yet representative clinical trial enrollment to evaluate these preventive measures. However, misinformation around the COVID-19 pandemic and general concerns about clinical trial participation in the U.S. hindered clinical trial enrollment. This study assessed awareness of, willingness to participate in, and enrollment in COVID-19 vaccine and treatment clinical trials in Texas. A quota sample of 1,089 Texas residents was collected online from June - July 2022. Respondents were asked if they were aware of, willing to participate in, and had enrolled in clinical trials for COVID-19 vaccines or treatments. Overall, 45.8% of respondents reported being aware of clinical trials for COVID-19 treatments or vaccines, but only 21.7% knew how to enroll and only 13.2% had enrolled in a COVID-19 clinical trial. Respondents with bachelor's or graduate degrees were more likely to be aware of clinical trials, more likely to have enrolled in trials, and more willing to participate in treatment trials. Women were less willing to participate and less likely to have enrolled in COVID-19 clinical trials than men. Respondents aged 55 years and older were more willing to participate, but less likely to have enrolled in COVID-19 clinical trials than 18-to-24-year-olds. Common reasons given for not participating in clinical trials included concerns that COVID-19 treatments may not be safe, government distrust, and uncertainty about what clinical trial participation would entail. Substantial progress is needed to build community awareness and increase enrollment in clinical trials.Item Demographic and Psychosocial Correlates of COVID-19 Vaccination Status among a Statewide Sample in Texas(MDPI, 2023-04-28) Luningham, Justin M.; Akpan, Idara N.; Taskin, Tanjila; Alkhatib, Sarah A.; Vishwanatha, Jamboor K.; Thompson, Erika L.The COVID-19 pandemic has been a global public health concern since early 2020 and has required local and state-level responses in the United States. There were several Food and Drug Administration (FDA) approved vaccines available for the prevention of COVID-19 as of August 2022, yet not all states have achieved high vaccination coverage. Texas is a particularly unique state with a history of opposing vaccination mandates, as well as a large and ethnically/racially diverse population. This study explored the demographic and psychosocial correlates of COVID-19 vaccinations among a statewide sample in Texas. A quota sample of 1089 individuals was surveyed online from June-July 2022. The primary outcome in this study was COVID-19 vaccination status (fully vaccinated, partially vaccinated, or unvaccinated) and included independent variables related to demographics, COVID-19 infection/vaccine attitudes and beliefs, and challenges related to the COVID-19 pandemic. Hispanic/Latinx individuals were more likely than non-Hispanic White individuals to be partially vaccinated as opposed to unvaccinated. Higher education levels and confidence that the FDA would ensure a safe COVID-19 vaccine were strongly associated with a higher likelihood of being fully vaccinated. In addition, some challenges brought on by the pandemic and concerns about becoming infected or infecting others were associated with a higher likelihood of being partially or fully vaccinated. These findings emphasize the need to further investigate the interaction between individual and contextual factors in improving COVID-19 vaccination rates, especially among vulnerable and disadvantaged populations.Item Development of methylene blue-loaded nanoparticles for glioblastoma treatment(2015-03) Castañeda-Gill, Jessica M.; Ranjan, Amalendu P.; Yang, Shaohua; Vishwanatha, Jamboor K.Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults over 45, resulting in an average survival of 15 months post-diagnosis and treatment. While recent research has provided essential information to aid diagnosis and treatment, GBM is known to cause relapse following traditional combinatorial regimens, necessitating the development of more effective and less toxic therapies. Methylene blue (MB), a dye with noted medicinal applications, has received recent consideration as a potential neurotherapeutic due to its ability to infiltrate the blood-brain barrier (BBB) and improve cellular processes within distinct brain cell compartments and types; however, one drawback is an increased administration to produce desired therapeutic effects, leading to excessive brain deposition and potential neurotoxicity. A method commonly used to enhance drug delivery while reducing unwanted side effects is via encapsulation in nanoparticles (NPs) composed of the biodegradable/biocompatible co-polymer, poly(lactic-co-glycolic) acid (PLGA). Based on our previous studies, we are developing a MB-loaded PLGA NP capable of permeating the BBB to treat GBM, to test our hypothesis that MB encapsulation into PLGA NPs will enhance accumulation in cancerous brain regions, resulting in reduced tumor size and prolonged survival. In this study, we formulated and characterized MB-loaded PLGA NPs, with a 3:1 molar ratio of sodium oleate to methylene blue at 5mg, based on particle size, drug loading, encapsulation efficiency, and release kinetics. Currently, we are establishing their in vitro effects in two different commercially-available GBM cell lines, according to their responses to commonly-used chemotherapeutics. Following loading of 5mg MB and their comparison to blank NPs, we obtained NP preparations in the range of 120-145nm, with encapsulation efficiencies from 25-40% and drug loading between 1-2%. Additionally, we have found that 50% of the MB initially added is released at 24 hours, and stays constant up to two weeks, demonstrating sustained drug release. In conclusion, based on studies that have demonstrated in vitro effects with MB at a minimum of 1μM (~0.3mg) and 150nm particles, our formulation should elicit comparable, if not better, results when treating GBM.Item EFFECT OF 4-HYDROXYNONENAL ON MIGRATION AND INVASION ENHANCER PROTEIN 1 (MIEN1) IN COLORECTAL CANCER(2014-03) Raychaudhuri, Urmimala; Vishwanatha, Jamboor K.MIEN1 could be a potential target for therapy in colorectal cancer. Purpose (a): Colorectal cancer (CRC) is the second leading cause of death in the United States. It is believed that the intestinal mucosa is constantly challenged with diet- and bacterial-derived oxidants and carcinogens. Chronic exposure of such challenging conditions may lead to the generation of reactive oxygen species (ROS). ROS initiate an autocatalytic chain of lipid peroxidation (LPO) of polyunsaturated fatty acids, resulting in the formation of large amounts of toxic electrophilic species and free radicals that may play important roles in various human diseases, including carcinogenesis. Consequently, even a minimal transient exposure of cells to ROS causes substantial lipid peroxidation, leading to a significant rise in the level of LPO end product, 4-hydroxynonenal (4-HNE), which is considered to be one of the most abundant cytotoxic aldehydes. HNE reacts not only with DNA but also with proteins and other molecules containing thiol and other nucleophilic groups and can alter the protein structure and functions. We have identified a novel protein called Migration and Invasion Enhancer protein 1 (MIEN1), is highly overexpressed in cancer cells and modulates the AKT activity as a membrane bound adaptor protein. Ectopic expression of MIEN1 activates Akt mediated downstream signaling through NF-kB pathway and induces the expression of several migratory and invasive proteins. However, 4-HNE has also been reported to induce the expression of various proteins involved in cell proliferation and migration. We hypothesize that 4-HNE mediated oxidative stress plays an important role in the etiology of colorectal cancer by modulating the expression and function of MIEN1. In the present studies, we have addressed this question by investigating the effect of 4-HNE on MIEN1 expression in colorectal cancer cell lines SW480 and HT29. Methods (b): Colorectal cancer cell lines, SW480 and HT29 were grown in RPMI-1640 medium containing 10% fetal bovine serum, in a humidified incubator at 37°C with 5% CO2. The toxicity of 4HNE in SW480 cells was determined by MTT assay. The effect of 4HNE on MIEN1 expression was determined by Western blotting in HT29. The effect of 4HNE on SW480 cell migration was examined by scratch wound assay. The LigandFit docking program available in the Accelrys molecular modeling software – Discovery studio, was used to carry out a docking study for the protein MIEN1 and substrate 4HNE. The 3D structure of the protein was obtained from PDB. Results (c): Our results demonstrated that exposure of 4HNE to SW480 cells is toxic. 4HNE concentrations ranging from 0 to 250 μM gradually decreased cell viability in SW480 cells corresponding to an IC50 value of 160 μM. Furthermore, our Western blot analysis demonstrated that treatment of 4HNE increased the expression of MIEN1 at the protein level in HT-29 cells. The scratch wound healing assay showed an increase in migration after treatment with low doses of 4HNE. The docking study produced 10 top scoring(dock score) poses. The poses indicate a possible interaction between the protein binding sites and the substrate (4HNE) including formation of hydrogen bonds between them. Conclusions (d): Together, these results suggest that 4HNE induced cell migration could be mediated via MIEN1.Item Effect of CRISPR MIEN1 knockout in metastatic breast cancer cells(2018-12) Van Treuren, Timothy; Vishwanatha, Jamboor K.; Basu, Alakananda; Basha, RiyazMigration and Invasion Enhancer 1 (MIEN1) is an oncogene which is involved in facilitating the migration and invasion of cancer cells through actin dynamics and gene expression. Increased MIEN1 expression in many types of tumors correlates with disease progression and metastatic propensity. The precise mechanism by which MIEN1 functions is yet to be understood. The goal of these studies is to progress toward determination of the mechanisms and genetic context in which MIEN1 functions contribute to cancer progression. It was hypothesized that Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) mediated knockout of MIEN1 in metastatic breast cancer cells would result in reduced migration and invasion. CRISPR genome editing effectively produced specific genomic deletions in the MIEN1 gene which led to the elimination of its expression in these breast cancer cells. Migration in MDA-MB-231 (231) MIEN1 knockout (MIEN1-KO) cells exhibited no difference when compared to parental 231, which was in contrast with previous siRNA studies. Signaling in several MIEN1-KO pools was inconsistent. Knocking out MIEN1 in 231 derivative cell lines showed few significant alterations in the growth, migration, invasion, signaling, despite significant changes in metabolism. However, re-expression of the MIEN1 protein containing a mutant immunoreceptor tyrosine-based activation motif (ITAM) domain resulted in significantly decreased invasion. This revealed that MIEN1-KO 231 derivative cells were susceptible to interference of compensatory mechanisms and demonstrates the importance of the migration and invasion pathways in which MIEN1 participates in breast cancer metastasis. These findings also suggest MIEN1 may still be a promising therapeutic target to inhibit metastasis if inhibitors can be developed which block ITAM function without affecting localization or expression.Item Effect of CRISPR MIEN1 Knockout in Metastatic Breast Cancer Cells(2018-12-01) Van Treuren, Timothy R.; Vishwanatha, Jamboor K.; Basu, Alakananda; Basha, RiyazMigration and Invasion Enhancer 1 (MIEN1) is an oncogene which is involved in facilitating the migration and invasion of cancer cells through actin dynamics and gene expression. Increased MIEN1 expression in many types of tumors correlates with disease progression and metastatic propensity. The precise mechanism by which MIEN1 functions is yet to be understood. The goal of these studies is to progress toward determination of the mechanisms and genetic context in which MIEN1 functions contribute to cancer progression. It was hypothesized that Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) mediated knockout of MIEN1 in metastatic breast cancer cells would result in reduced migration and invasion. CRISPR genome editing effectively produced specific genomic deletions in the MIEN1 gene which led to the elimination of its expression in these breast cancer cells. Migration in MDA-MB-231 (231) MIEN1 knockout (MIEN1-KO) cells exhibited no difference when compared to parental 231, which was in contrast with previous siRNA studies. Signaling in several MIEN1-KO pools was inconsistent. Knocking out MIEN1 in 231 derivative cell lines showed few significant alterations in the growth, migration, invasion, signaling, despite significant changes in metabolism. However, re-expression of the MIEN1 protein containing a mutant immunoreceptor tyrosine-based activation motif (ITAM) domain resulted in significantly decreased invasion. This revealed that MIEN1-KO 231 derivative cells were susceptible to interference of compensatory mechanisms and demonstrates the importance of the migration and invasion pathways in which MIEN1 participates in breast cancer metastasis. These findings also suggest MIEN1 may still be a promising therapeutic target to inhibit metastasis if inhibitors can be developed which block ITAM function without affecting localization or expression.Item The Effect of Trusted News Sources on the Confidence in the Safety of COVID-19 Vaccination(2023) Alkhatib, Sarah A.; Luningham, Justin M.; Akpan, Idara N.; Taskin, Tanjila; Vishwanatha, Jamboor K.; Thompson, Erika L.Purpose: COVID-19 vaccination prevents severe disease manifestations; yet uptake has been suboptimal. Confidence in the safety and efficacy of the vaccine influences COVID-19 vaccination decisions. Exposure to information from a trusted news source can impact perceptions and may contribute to vaccine decisions. This study assessed the association between trusted news sources and confidence in the safety of COVID-19 vaccination among Texas adults. Methods: Participants were recruited through an online panel using quota sampling based on the racial and ethnic distribution of Texas in July 2022 (n=1089). The primary predictor variable was self-reported trusted news sources for COVID-19 related news (16 options), in which respondents were asked to endorse any news source they trusted, with options ranging from print media to cable news to local news. The outcome was confidence in the safety of the COVID-19 vaccine (not at all confident to very confident). Multinomial regression analyses were conducted to model confidence in COVID-19 vaccination and trusted news sources while controlling for education, age, gender, and self-reported race. Results: Through an initial descriptive analysis, Fox News, local cable TV programs, and news broadcasting from one’s home abroad were associated with lower confidence levels. After grouping those three sources as "top news sources” and assessing their effect on confidence through a multinomial model, it was found that individuals who trusted those top sources were significantly less likely to endorse "somewhat confident” (OR=0.59, 95%CI 0.4-0.89) or "very confident” (OR=0.41, 95%CI 0.27- 0.62) compared to being "not at all confident” in the safety of the COVID-19 vaccine. Conclusions: Study findings show that some trusted news sources contributed to participants having less confidence or no confidence in the safety of the COVID-19 vaccine. Public health initiatives should consider how to address vaccine confidence among the public given the diversity of information sources people rely on.Item Enhancing research careers: an example of a US national diversity-focused, grant-writing training and coaching experiment(BioMed Central Ltd., 2017-12-04) Jones, Harlan P.; McGee, Richard; Weber-Main, Anne Marie; Buchwald, Dedra S.; Manson, Spero M.; Vishwanatha, Jamboor K.; Okuyemi, Kolawole S.BACKGROUND AND PURPOSE: Preparing a successful research proposal is one of the most complex skills required of professional scientists, yet this skill is rarely if ever, taught. A major goal of the National Research Mentoring Network (NRMN) in the United States (U.S.) is to support the professional advancement of postdoctoral fellows and junior faculty from diverse populations by offering intensive coaching in the development of grant proposals early in their careers. This article highlights the National Institutes of Health's (NIH) NRMN initiative to prepare diverse constituencies of early-stage biomedicine scientists for research careers by implementation of an evidence-based nationwide program of comprehensive grant writing and professional development. PROGRAM AND KEY HIGHLIGHTS: NRMN delivers four unique but complementary coaching models: the Proposal Preparation Program from the University of Minnesota (UMN); Grantwriters Coaching Groups from Northwestern University (NU); Grantwriting Uncovered: Maximizing Strategies, Help, Opportunities, Experiences from the University of Colorado Anschutz Medical Campus (UC) and Washington State University (WSU); and Steps Towards Academic Research from the University of North Texas Health Science Center (UNTHSC). Because these programs cater to scientists at different career stages, rather than employ a single approach, each is uniquely tailored to test its efficacy at the national level. The first two models prioritize scientists with reasonably well-developed research projects who are ready to write proposals for specific NIH research competitions. The other two models target postdoctoral fellows and early-career faculty who need more extensive guidance in proposal development plans. To achieve scalability, all programs also recruit faculty as Coaches-in-Training to learn approaches and acquire particular group facilitation skills required by each model. IMPLICATIONS: These efforts exemplify NRMN's potential to enhance the career development of diverse trainees on a national scale, building research skills, competitiveness for obtaining faculty positions and capacities that will result in high quality research proposals from a diverse pool of applicants, thereby advancing innovations in science and diversifying the U.S. biomedical workforce.Item Exosomal AnnexinA2 promotes angiogenesis and breast cancer metastasis(2015-03) Maji, Sayantan; Akopova, Irina; Nguyen, Phung M.; Hare, Richard J.; Gryczynski, Ignacy; Vishwanatha, Jamboor K.Purpose: Early detection of cancer using circulating biomarkers is a realistic possibility with the discovery of exosomes. Cells under both physiological and pathological conditions secrete a wide array of membranous vesicles containing specific protein and RNA signatures. Exosomes, which are 40-100nm in size, comprise a major portion of these vesicles. Annexin A2 (AnxA2) is a 36 KD Ca+2 dependent phospholipid-binding protein up-regulated in many cancer types and implicated in promoting tumorigenesis and angiogenesis. Cancer cells have been shown to secrete more AnxA2 and it is also highly expressed in the exosomes; but the function of exosomal AnxA2 (exo-AnxA2) has never been studied. We hypothesize that exo-AnxA2 promotes angiogenesis and breast cancer metastasis and we propose to explore this in our study. Methods: Exosomes were isolated from MCF10A progression model for progression studies. For the metastasis studies MDA-MB-231 and its organ specific metastatic variants MDA-MB-831 (brain met) and MDA-MB-4175 (lung met) were used. They were characterized via Western blotting, particle size analyzer, transmission Electron Microscopy (TEM) and Atomic Force Microscopy (AFM). In vitro and in vivo angiogenesis assays were performed to study the role of exo-AnxA2 in angiogenesis. Exo-AnxA2 was either inhibited by use of LCKLSL inhibitory peptide or knocked down in these studies. Tail vein and intracardiac injection metastasis models were used after priming the animals with exosomes to study the role of exo-AnxA2 in breast cancer metastasis. Results: Characterization of exo-AnxA2 by Western blot and AFM revealed that it is highly expressed in cancer exosomes than exosomes from normal cells. In vitro and in vivo angiogenesis studies revealed exo-AnxA2 to be a potent inducer of angiogenesis and inhibition of exo-AnxA2 significantly inhibits angiogenesis (~3 fold and ~5 fold decrease with LCKLSL vs. control in endothelial assay and matrigel plug assay respectively). Tail vein and intracardiac metastasis models showed that exo-AnxA2 promotes lung and brain metastasis. Knockdown of exo-AnxA2 showed ~2 fold decrease in lung and brain metastasis vs. control respectively, as evident from bioluminescence imaging. Conclusion: We found that exo-AnxA2 correlates positively with breast cancer progression. Furthermore, we found that exo-AnxA2 is a potent inducer of angiogenesis and breast cancer metastasis indicating a possible role of exo-AnxA2 in tumor- microenvironment signaling and cancer progression.Item Factors associated with COVID-19-related mental health among Asian Indians in the United States(Elsevier B.V., 2023-01-11) Ikram, Mohammad; Shaikh, Nazneen F.; Siddiqui, Zasim A.; Dwibedi, Nilanjana; Misra, Ranjita; Vishwanatha, Jamboor K.; Sambamoorthi, UshaBACKGROUND: In the United States, the COVID-19 pandemic has caused increased mental health symptoms and mental illness. Specific subgroups such as Asian Indians in the US have also been subject to additional stressors due to unprecedented loss of lives in their home country and increased Asian hate due to the misperception that Asians are to be blamed for the spread of the SARS-CoV-2. OBJECTIVE: We examined the various factors including discrimination associated with COVID-19-related mental health symptoms among Asian Indians. METHODS: We administered an online survey between May 2021 and July 2021 using convenient and snowball sampling methods to recruit Asian Indian adults (age > 18 years, N = 289). The survey included questions on mental health and the experience with unfair treatment in day-to-day life. Descriptive analysis and logistic regressions were performed. RESULTS: Overall, 46.0% reported feeling down, depressed, or lonely and feeling nervous, tense, or worried due to the COVID-19 pandemic; 90.0% had received at least one dose of vaccination and 74.7% reported some form of discrimination. In the fully-adjusted logistic regression, age (AOR = 0.95; 95%CI- 0.92, 0.97;p < 0.01) and general health (AOR=0.84; 95%CI- 0.73, 0.97; p < 0.015) were negatively associated with mental health symptoms. Participants who experienced discrimination were more likely (AOR=1.26; 95%CI- 1.08, 1.46; p < 0.01) to report mental health symptoms. CONCLUSION: In this highly vaccinated group of Asian Indians discriminatory behaviors were associated with mental health symptoms suggesting the need for novel institutional level policy responses to reduce anti-Asian racism.