Research Award Program Research Articles

Cross-Sectional Variation of Option-Implied Volatility Skew

The stock option-implied volatility skew reflects both the structural risk characteristics of the underlying company and the short-term information flow about the stock price movement. This paper builds a semistructural, cross-sectional option pricing model to separate the structural risk contributions from the information flow. The model identifies two structural risk sources that contribute to the cross-sectional variation of the skew: the company’s business cyclicality and its default risk. The model can explain as much as 44% of the cross-sectional variation in implied volatility skew and is particularly informative during and after recessions. The remaining skew variation reflects mainly short-term information flow and can be used to construct stock portfolios with much better investment performance and without hidden structural risk exposures. This paper was accepted by Agostino Capponi, finance. Funding: L. Wu gratefully acknowledges support by a grant from the City University of New York PSC-CUNY Research Award Program. Supplemental Material: The online appendix and data are available at https://doi.org/10.1287/mnsc.2023.4872 .

Investigating the Impact of Hydrocephalus on Epiplexus Cells of the Choroid Plexus

Hydrocephalus is characterized by accumulation of cerebrospinal fluid (CSF) in the brain, resulting from over-production, under-absorption, or blockage of the flow of CSF. Current treatments are limited to surgically placed shunts that often require follow-up procedures. The goal of this research group is to understand the mechanisms of hydrocephalus with the aim of finding effective pharmacological treatments. The animal model used exhibits a mutation in the transmembrane 67 (Tmem67) gene. Homozygous recessive animals present with ventriculomegaly at birth and severe hydrocephalus by postnatal day 15 (P15). Experiments in this study compare tissue obtained from wildtype animals and homozygous mutant animals. The choroid plexus (CP) is the main producer of CSF, making it a target structure in hydrocephalus research. It is a branched tissue that includes a stroma with fenestrated capillaries, wrapped by a tightly regulated barrier epithelium. Choroid plexus epithelial cells (CPe) are multiciliated cells that strictly regulate the osmotic balance of CSF. Epiplexus cells are poorly understood immune cells localized on the apical (CSF-facing) side of the CPe. These cells are believed to share functional characteristics with microglia and macrophages. Microglia are the innate immune cells of the central nervous system; a specialized type of glial cell that interact with neurons, and aid in the structure and immunosurveillance of the brain. Macrophages originate from monocytes and reside in most tissues of the body, responding to cellular signals of injury and infection. Macrophages can be found free-roaming in tissues or associated to epithelial cells, known as border-associated macrophages (BAMs). Due to their association with the CPe, epiplexus cells are considered BAMs. Beyond their apparent similarities to other immune cells, little else is known about the properties and functions of epiplexus cells or their interaction with the CPe or CSF, nor if they play a role in the development of hydrocephalus. This study furthers the understanding of epiplexus cells, specifically how they impact, or are impacted by hydrocephalus. Rat pups (P15) were terminally anesthetized, perfused with 1x PBS, and fixed with 4% PFA. In the lateral ventricle CP, we have visualized CPe-associated cells with morphology consistent with epiplexus cells. These cells are labeled with microglial/macrophage/epiplexus cell markers: allograft inflammatory factor1/ionized calcium-binding adaptor molecule 1 (Iba1), macrosialin (CD68), and mannose receptor type C-1 lectin (CD206). Iba1 is thought to play a role in restructuring the actin cytoskeleton in cells of monocyte lineage (macrophages and microglia), resulting in morphological differences across levels of activation. CD68 is a transmembrane protein commonly expressed in macrophages and monocytes, supposedly related to the oxidized low-density lipoprotein (OxLDL) pathway, which is involved in exacerbating oxidative stress. CD206 is a transmembrane protein that may aid in the roles of detecting pathogens, removing proteins, and sending waste through lymphatic system. Confocal analysis combining cell morphology and immunohistochemical labeling will be used to quantify CPe-associated epiplexus cells in wildtype and hydrocephalic animals. The results will be substantiated using quantitative real-time PCR. Funding: United States Department of Defense Congressionally Directed Medical Research Program Award; Hydrocephalus Association. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

TRPV4 Antagonist Treatment of Hydrocephalus Causes Cell and Molecular Changes in Multiple Brain Cell Types

Hydrocephalus is defined by cerebrospinal fluid (CSF) accumulation in the brain’s ventricles causing ventriculomegaly, an imbalance in CSF secretion, flow, and/or absorption. Treatments for hydrocephalus include invasive surgical procedures, which commonly fail throughout a patient’s life. Transient receptor potential vanilloid member 4 (TRPV4) is a mechanosensitive cation channel implicated in osmotic regulation. Our laboratory found that a TRPV4 antagonist, RN1734, ameliorates hydrocephalus in a rat model of congenital hydrocephalus. It was hypothesized that treatment slowed production of CSF by targeting choroid plexus (CP), a contiguous layer of tightly regulated epithelial cells, that is responsible for CSF production. However, hydrocephalus pathology can have various effects on the brain. This study focuses on changes occurring in CP epithelial cells (CPe) and glia, which have roles in brain-barrier maintenance and brain fluid/electrolyte regulation. Interestingly, CPe and glia contain many of the same channels and transporters, including TRPV4, that are important in regulating both CSF and brain interstitial fluid. These cells contain aquaporins (AQP), AQP1 in CPe, and AQP4 in glia. AQPs have been implicated in diseases associated with fluid regulation and may interact with TRPV4. We hypothesize that TRPV4 and AQPs are important in hydrocephalus pathology, and that changes may be attenuated with RN1734 treatment. A point mutation in TMEM67 results in hydrocephalus (hydro). Changes in cell morphology, gene transcription, protein expression, localization of TRPV4, AQP1, and AQP4 were observed. Postnatal day 15 (P15) untreated (unt) and RN1734-treated (RN;2mg/ml DMSO; 4mg/kg intraperitoneal injection daily P7-P14) wildtype and mutant ( Tmem67−/−) rats were utilized in this study. CPe and periventricular (PV) glia were visualized using fluorescent immunohistochemistry (IHC) of GFAP and a plasma membrane marker. Cell volume of CPe’s and process morphology of glia were altered in hydro rats. RN returned these changes to relatively normal in CPe, but not glia. Next, we used IHC to examine localization of TRPV4 and the AQPs. Increased immunoreactivity of TRPV4 and AQP1 was observed in CPe of unt and RN Tmem67−/− rats. AQP4 and TRPV4 immunoreactivity increased in the subventricular zone and PV white matter of hydro rats. TRPV4 immunoreactivity, but not AQP4, was normalized with RN. We then utilized RTqPCR to inspect transcriptional changes. The data showed increased AQP1 and decreased TRPV4 mRNA in PV cortex and CP of unt and RN Tmem67−/− rats. Previous data from our laboratory found no change in TRPV4 protein in unt Tmem67−/− CPe, but we were curious about AQP expression. The amount of AQP1 protein was increased in the CP of unt and RN Tmem67−/− rats. In PV cortex, AQP4, but not TRPV4, protein was increased in hydro rats. With RN, AQP4 protein was not normalized. In summary, TRPV4 changes in the Tmem67−/− rats returned to normal with RN, glial morphology and AQP changes did not. This may indicate residual inflammation in the Tmem67−/− rats, which we aim to address in future studies. To further elucidate TRPV4’s role in hydrocephalus pathology, future studies will explore post-translational and membrane localization changes. Overall, investigating cell and molecular changes that occur in hydrocephalus will help find better pharmacological targets for this disease. Funding: United States Department of Defense Congressionally Directed Medical Research Program Award; Hydrocephalus Association. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Speech for Intelligent cognition change tracking and DEtection of AD (SIDE‐AD) Research Program: SIDE‐AD

AbstractBackgroundThere is emerging evidence that speech could be a potential indicator and manifestation of early Alzheimer’s disease (AD) pathology. Symptomatic AD may change an individual’s language, especially certain elements in speech such as elaboration and attribution. Therefore, the University of Edinburgh and Sony Group Corporation are collaborating within the Sony Research Award Program to create the SIDE‐AD study which aims to develop digital speech‐based biomarkers technology for AD. The study is designed to assess disease status in the pre‐dementia stages in a real‐world at‐risk population in Brain Health Services.MethodSIDE‐AD is an observational longitudinal study developing speech biomarkers for use in neurodegenerative disease. Participants are recruited from Brain Health Services in Scotland with data collection starting in spring 2023. In the context of the SIDE‐AD study, we have developed a secured online data collection infrastructure for collecting voice data by prompting individuals to record a voice sample talking about their brain health. Individuals are asked to rate their mood, anxiety and apathy. The baseline speech biomarkers will be compared to the follow‐up visit’s speech recordings as well as to other variables from routinely collected health care data. The SIDE‐AD study will employ signal processing and machine learning technologies to automate the assessment of the respondents’ speech.ResultThe objective of the SIDE‐AD study is to recruit participants (n = 150) across the AD spectrum. The developed infrastructure will be used to collect data of participants with an objective of analysing baseline speech markers in relation to self‐reported mood, anxiety and apathy as well as clinical outcomes from patients’ health care records.ConclusionThe SIDE‐AD study is carrying out research and development of a speech‐based application recording data on individuals’ self‐perceived brain health. A secured online data collection infrastructure is developed to collect speech data with an objective to analyse speech data in combination with routinely available clinical data in Brain Health Clinics, enabling real world validation of speech‐based digital biomarkers for the early detection of AD. We plan to use automatic speech recognition and acoustic patterns for the automatic assessment of brain health status relevant to emergent AD‐specific speech biomarkers.

Combined GIP receptor and GLP1 receptor agonism attenuates NAFLD in male APOE∗3-Leiden.CETP mice.

Combined glucose-dependent insulinotropic polypeptide receptor (GIPR) and glucagon-like peptide-1 receptor (GLP1R) agonism is superior to single GLP1R agonism with respect to glycemic control and weight loss in obese patients with or without type 2 diabetes. As insulin resistance and obesity are strong risk factors for nonalcoholic fatty liver disease (NAFLD), in the current study we investigated the effects of combined GIPR/GLP1R agonism on NAFLD development. Male APOE∗3-Leiden.CETP mice, a humanized model for diabetic dyslipidemia and NAFLD when fed a high-fat high-cholesterol diet, received subcutaneous injections with either vehicle, a GIPR agonist, a GLP1R agonist, or both agonists combined every other day. GIPR and GLP1R agonism reduced body weight and additively lowered fasting plasma levels of glucose, triglycerides and total cholesterol. Strikingly, we report an additive reduction in hepatic steatosis as evidenced by lower hepatic lipid content and NAFLD scores. Underlying the lipid-lowering effects were a reduced food intake and intestinal lipid absorption and an increased uptake of glucose and triglyceride-derived fatty acids by energy-combusting brown adipose tissue. Combined GIPR/GLP1R agonism also attenuated hepatic inflammation as evidenced by a decreased number of monocyte-derived Kupffer cells and a reduced expression of inflammatory markers. Together, the reduced hepatic steatosis and inflammation coincided with lowered markers of liver injury. We interpretate that GIPR and GLP1R agonism additively attenuate hepatic steatosis, lower hepatic inflammation, ameliorate liver injury, together preventing NAFLD development in humanized APOE∗3-Leiden.CETP mice. We anticipate that combined GIPR/GLP1R agonism is a promising strategy to attenuate NAFLD progression in humans. This work was supported by a grant from the Netherlands CardioVascular Research Initiative: the Dutch Heart Foundation, Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences [CVON-GENIUS-II] to P.C.N.R., a Lilly Research Award Program [LRAP] Award to P.C.N.R. and S.K., a Dutch Heart Foundation [2017T016] grant to S.K., and an NWO-VENI grant [09150161910073] to M.R.B.; J.F.D.B. is supported by the Nutrition and Health initiative of the University of Groningen; Z.Y. is supported by a full-time PhD scholarship from the China Scholarship Council (201806850094 to Z.Y.).

Longitudinal analysis of the LA-SPARTA cohort of high-risk individuals after SARS-CoV-2 infection and vaccination

Abstract Questions remain regarding correlates of risk and immune protection against SARS-CoV-2 infection and vaccination response. Thus, we prospectively enrolled 200 participants with a high risk of SARS-CoV-2 occupational exposure. Longitudinal analysis of participant exposure risks, vaccination/infection status, and symptoms were assessed at 3, 6, and 12 months, with blood and saliva collection. Quantitative ELISA assay was used to assess the serological response to the SARS-CoV-2 spike holoprotein (S), S-receptor binding domain (RBD) and nucleocapsid protein (NP). 40 of 200 (20%) participants were found to be infected via serology. Equal incidence of infection was found in healthcare and non-healthcare occupations. In those infected, only 79.5% seroconverted for NP, and 11.5% were unaware they had been infected. The antibody response to S was greater than to RBD. Additionally, Hispanic ethnicity was associated with 2-fold greater incidence of infection despite vaccination. Booster vaccination significantly increased the avidity of RBD-specific responses to variant spike proteins. No differences in avidity were found if a participant was boosted via vaccination or natural infection. Collectively, our results indicate that antibody quality and holoprotein response may reveal meaningful determinants of protection beyond RBD antibody titer alone. Additionally, determinants of infection risk may include race/ethnicity, and remain independent of comorbidity and humoral response in this cohort. This study received funding from Center for Influenza Vaccine Research for High Risk Populations- CIVIC 75N93019C00052 (EFR), Lundquist Institute Institutional Funds (MRY), UCLA W. M. Keck Foundation COVID 19 Research Award Program (EFR), and the Community Engagement Alliance Against COVID-19 (CEAL) (MRY, EFR). The project described was supported by the National Center for Advancing Translational Sciences through the UCLA CTSI Grant UL1TR001881. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH or other funding sources. MMJ is supported by the NIH grants TL 1 DK132768 and U2C DK129496.

Unveiling the antiviral activity of interferon epsilon in respiratory infections

Abstract Type I interferons (IFN-I), including IFN-α and IFN-β, have demonstrated efficacy in treatments against some viral infections but with a considerable amount of undesirable side effects. IFN-ɛ, a recently discovered member of the IFN-I family, has been found to be expressed constitutively in several tissues, including the lung. However, the antiviral effect of IFN-ɛ against respiratory viral infections is largely unknown. Human respiratory syncytial virus (RSV) is the most important agent responsible for acute lower respiratory tract diseases in infants worldwide. Each year in the United States, RSV leads to approximately 2.1 million outpatient (non-hospitalization) visits among children younger than five years old, with a considerable number of hospitalizations among children younger than five years old and adults 65 years and older. Currently, no vaccine is available to prevent this respiratory tract infections, and new strategies are necessary to treat patients. Therefore, in this study, we aimed to investigate the antiviral effect of IFN-ɛ against respiratory viruses, such as RSV. Using a human epithelial cell line, we assessed the expression, susceptibility, and antiviral effect of IFN-ɛ on RSV infection. Our results demonstrate that RSV is capable of inducing IFN-ɛ. Moreover, RSV is susceptible to the antiviral activity of the rhIFN-ɛ, reducing the number of infected cells as well as the viral replication. That effect is due to the induction of the expression of ISGs with a low inflammatory profile. Overall, our findings demonstrate that RSV is susceptible to the effect of IFN-ɛ, suggesting a potential therapeutic effect of this novel member of the IFN-I family to treat respiratory viral infections. This work was funded in part by a Louisiana State University School of Veterinary Medicine Competitive Research Program Award

The impact of the complement receptors C3aR1 and C5aR1 on the progression of melanoma

Abstract Melanoma is an aggressive skin cancer that develops from the malignant transformation of pigment-producing skin cells, melanocytes. The incidence of cutaneous melanoma is remarkably high, with an estimated number of new cases in the United States in 2022 close to 100,000 patients. Several previous reports pointed out the effect of the complement system in the progression of melanoma, although the precise mechanism is largely unknown. The complement system is a crucial component of innate immunity, and it also has a significant role in adaptive immunity regulating the function of immune cells. The complement receptors C3aR1 and C5aR1 are present on the surface of various immune cells. Anaphylatoxins (C3a and C5a) generated by complement activation bind to their respective receptors, C3aR1 and C5aR1, suppress the antitumor function of immune cells and promote migration and activity of immunosuppressive cells in the tumor microenvironment. Hence, C3aR1 and C5aR1 act as immune checkpoint receptors. To identify the precise role of the complement receptors in melanoma, we challenged C3 −/−, C3aR1 −/−, and C5aR1 −/−mice with the B16F10 murine melanoma cells. We showed that the deficiency of these molecules substantially delayed tumor growth and promoted an antitumorigenic immune response. The results of this study indicate the distinct role of complement receptor signaling on melanoma growth and suggest a novel immunotherapeutic approach. Developmental Research Program Award from the MD Anderson Melanoma SPORE (P50CA221703-04)

Abstract LB039: Ovarian cancer cell-derived exosomal UCA1 reprograms glucose metabolism in stromal fibroblasts

Abstract Tumor progression is stringently regulated by the co-ordinated signaling between the tumor cells and the tumor microenvironment (TME) through diverse autocrine, paracrine and exocrine signaling mechanisms. Recent studies have shown that the exosomes that play a crucial role in the transfer of oncogenic macromolecules and oncometabolites to different cellular components of the TME can be targeted for therapy. Therefore, with the focus on identifying the exosomal long non-coding RNA (lncRNA) that can be therapeutically targeted, we investigated the profile of ovarian cancer cell derived lncRNAs and their functional role in ovarian cancer pathophysiology. Analysis of exosomes derived from a panel of high grade serous ovarian cancer cell lines indicated that UCA1 (Urothelial Cancer Associated 1) is the most abundantly packaged lncRNA in the exosomes. Similarly, exosomes from patient-derived ovarian cancer cells exhibited higher levels of UCA1. Ascites from ovarian cancer patients also indicated high levels of UCA1 in the ascites-derived exosomes. Results using PKH67-labelled ovarian cancer cell-derived exosomes and MRC5 fibroblast cell line indicated the exosomal transfer of UCA1 to the fibroblasts. Since paracrine signaling has been shown to induce metabolic reprogramming of peritumoral fibroblasts, we interrogated whether exosomal transfer of UCA1 could reprogram the glucose metabolism in the fibroblasts. Results from the Agilent Seahorse glycolytic stress assay indicate that the exosomal UCA1 promotes reprogramming of glucose metabolism in fibroblast cells while depleting of UCA1 in the exosomes failed to induce such metabolic reprogramming. Results from the analysis of the key glycolytic enzymes in the stromal fibroblasts, post-exosomal UCA1 uptake also corroborate this conclusion. Thus our results provide primary evidence that the exosomal-UCA1 induces pro-tumorigenic metabolic reprogramming in peri-tumoral fibroblasts. Together with the known oncogenic role of UCA1 in many different cancer cells, our findings indicate that targeting UCA1 could form a productive precision cancer strategy in ovarian cancer, targeting both the tumor cells and tumor stromal cells. This research was supported by the Department of Defense Ovarian Cancer Research Program Award W81XWH-18-1-0066, W81XWH-22-1-0415, the National Institute of General Medical Sciences grant P20 GM103639 and The National Cancer Institute of the National Institutes of Health grant P30 CA225520. Citation Format: Revathy Nadhan, Ji Hee Ha, Muralidharan Jayaraman, Srishti Kashyap, Danny N. Dhanasekaran. Ovarian cancer cell-derived exosomal UCA1 reprograms glucose metabolism in stromal fibroblasts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB039.

Distinct Patterns in the Reconstitutions of Treg and T-Helper Subsets after Autologous Hematopoietic Stem Cell Transplantation in Juvenile-Onset Systemic Sclerosis Patients

Systemic sclerosis (SSc) is a refractory autoimmune disease characterized by diffuse fibrosis and vasculopathy affecting the skin, joints, and internal organs. Most immunosuppressive drugs have failed to show long-term benefit in randomized trials. In recent years, autologous hematopoietic stem cell transplantation (ASCT) has been performed to treat multiple sclerosis and SSc, however, the reconstitutions of immune components post-ASCT in SSc have not been fully described. Subjects and Methods: We tested a total of 13 samples from 4 consenting SSc patients (3 females, 1 male) receiving IRB-approved immunoablation (TBI 600cGy) with lung shielding and Cytoxan (n=3) or Thiotepa (n=1) +/- Alemtuzumab followed by autologous CD34+ selected, cryopreserved peripheral blood hematopoietic stem cells transplantation. Their median age was 18.5-years-old (range 15.2- to 21.4-years-old). Peripheral blood samples were collected at baseline pre-ASCT, and 3, 6, 12, 24-months post-ASCT. By flow cytometry, we examined lymphocyte and monocyte subset recovery with additional focus on the FOXP3+Treg and T-helper subset reconstitution. Results and Discussion: All 4 subjects experienced significant improvement in several clinical and quality of life parameters, especially in their skin disease evident within weeks after ASCT. T-cell lymphopenia was profound at 3-months; however, the recovery was evident by 6-months reaching pre-ASCT levels by 2-years post-ASCT (Fig 1A). Natural killer (NK) lymphocytes and monocytes attained the normal values by 3-months and after (Fig 1A). The FOXP3+Tregs that appeared post-ASCT displayed distinct features. There was an increased percentage of FOXP3+Tregs in total CD4+T cells during the early stage (3- to 6-months) post-ASCT (Fig 1B), leading to an increase in the ratio of Treg/Tcon (Fig 1B). The higher proportions of proliferating Tregs as measured by Ki67+FOXP3+CD4+/CD25+/CD127- T cells (Fig 1C) likely led to their increased numbers. This trend was sustained until the last evaluation at 2-years post-ASCT (Fig 1C). In contrast, CD4+Tcons showed a relatively lower proliferation rate, leading to a higher ratio of Ki67+Treg/Ki67+Tcon compared to pre-ASCT values (Fig 1C). Notably, there was an increased proportion of Tregs expressing the skin homing marker cutaneous lymphocyte-associated antigen (CLA) at 3-months lasting until the last test at 2-years post-ASCT (Fig 1D), whereas CLA+/CD4+Tcons were less plentiful. The reconstitutions of T-helper (Th) subsets were also examined. Due to lymphopenia at the early stage post-ASCT, Th subsets were identified according to their chemokine receptor expression (Mahnke et alCytometry 2013;83(5):439-440). Gradually decreased numbers of Th2, Th9, and Th17 were observed in the peripheral pool (Fig 1E) contributing to a decline of their proportions in total CD4+T (Fig 1F). Nevertheless, absolute number of Th1 and Th22 cells returned to levels seen pre-ASCT without any recurrence or flare of SSc symptoms. The CD45RO+ memory phenotype was dominant during the early stages post-ASCT in all subsets (data not shown). Summary: Successful reconstitution of lymphocytes and monocytes post-ASCT was observed in available SSc patients. Post-ASCT circulating FOXP3+Tregs displayed distinct features where the increased number led to an elevated Treg/Tcon ratio in the peripheral pool. Amongst Tregs, the skin homing CLA+ subset was further enriched correlating with improved skin scores in all patients. Improved peripheral proliferation and increased expression of CLA+ Tregs may provide protection from skin disease recurrence despite parallel Tcon reconstitution. The decrease of the Th2/Th9/Th17 subsets in the peripheral pool post-ASCT possibly contributes to the clinically less autoimmune phenotype. These results need to be validated in future patients. Acknowledgements: We thank all participating patients, nurses, physicians, and scientists in this study for their contributions. The study was supported by the "Idea Development Award" from the Department of Defense - Scleroderma Research Program (Award number W81XWH-21-1-0782 and W81XWH-21-1-0783). Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Letter from the Editor

Produced by the Division of Learning and Teaching Support and Innovation at the University of Victoria, The Arbutus Review was created to showcase the articles, projects, and installations that result from the Jamie Cassels Undergraduate Research Award (JCURA) program. JCURA was instituted in 2009 as the Undergraduate Research Scholarship program by Jamie Cassels, then Vice-President Academic and Provost and President from 2013–2020. The award program was designed to support and create truly formative research experiences for exceptional undergraduate students. The Division of Learning and Teaching Support and Innovation administers the award nomination process on behalf of the Provost’s Office. In addition to submissions that were the result of JCURA research, The Arbutus Review also publishes other exceptional work from students in departments across campus.

Genome Sequence Resource for Strains of Pseudomonas syringae Phylogroup 2b and P. viridiflava Phylogroup 7a Causing Bacterial Stem Blight of Alfalfa.

Genome Sequence Resource for Strains of Pseudomonas syringae Phylogroup 2b and P. viridiflava Phylogroup 7a Causing Bacterial Stem Blight of Alfalfa.

Epithelial-derived oxysterol production tunes intestinal IgA secretion against commensals and enteric pathogen in tissue

Abstract Immunoglobulin A (IgA) secretion by plasma cells (PCs), terminally differentiated B cells residing in the intestinal lamina propria, assures microbiome homeostasis and protects the host against enteric infections. However, whether exposure to diet-derived and commensal-derived signals instruct tissue resident PCs effector function and dynamically shape IgA immune responses at the mucosal barrier remain largely uninvestigated. Here, we demonstrated that intestinal epithelial cells (IECs) integrate luminal input to produce 7α,25-dihydroxycholesterol (7α,25-HC), a cholesterol metabolite (oxysterol) with immunomodulatory functions. In IECs, both cholesterol uptake via NPC1L1 and commensal recognition via MyD88 controlled oxysterol production. Mice lacking the oxysterol enzyme CH25H specifically in IECs abolished 7α,25-HC production and allowed to study oxysterol generation and activity in the small intestine. Inability of IECs to generate 7α,25-HC enhanced IgA secretion by PCs in the gut, suggesting that oxysterol negatively regulate humoral response at the mucosal barriers. Mechanistically, we showed that intestinal PCs sensed 7α,25-HC via the chemoattractant receptor GPR183 to position in the lamina propria tissue. This IEC-PC axis was rapidly modulated by cholesterol dietary content and tuned Salmonella-specific IgA response. Our finding revealed a new mechanism linking dietary cholesterol and humoral immune responses centered around PC localization for efficient mucosal protection. Supported by grants from Kenneth Rainin Foundation, Innovator Award, Charles H. Hood Foundation Child Health Research Awards Program, The Leukemia and Lymphoma Society New Idea Award and the Multiple Myeloma Research Fellowship, NIH ( AI40098 ) and The American Association of Immunologists Careers in Immunology Fellowship Program.

PD01-12 PAN-CANCER CHARACTERIZATION OF THE TUMOR MICROENVIRONMENT REVEALS STROMA AND IMMUNE ENRICHED SUBTYPE WITH DISTINCT MOLECULAR, CLINICAL, AND PROGNOSTIC FEATURES IN DIFFERENT GENITOURINARY CANCERS

You have accessJournal of UrologyCME1 May 2022PD01-12 PAN-CANCER CHARACTERIZATION OF THE TUMOR MICROENVIRONMENT REVEALS STROMA AND IMMUNE ENRICHED SUBTYPE WITH DISTINCT MOLECULAR, CLINICAL, AND PROGNOSTIC FEATURES IN DIFFERENT GENITOURINARY CANCERS Mark Farha, Brittney Cotta, Srinivas Nallandhighal, Kevin Hu, Steven Goldenthal, Aaron Udager, and Simpa Salami Mark FarhaMark Farha More articles by this author , Brittney CottaBrittney Cotta More articles by this author , Srinivas NallandhighalSrinivas Nallandhighal More articles by this author , Kevin HuKevin Hu More articles by this author , Steven GoldenthalSteven Goldenthal More articles by this author , Aaron UdagerAaron Udager More articles by this author , and Simpa SalamiSimpa Salami More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002516.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Transcriptomic data driven tumor microenvironment (TME) deconvolution tools have enabled robust analyses correlating TME subtypes with prognosis and therapeutic response. Here, we performed a pan-cancer analysis of tumors included in the TCGA to assess the impact of the TME on oncological outcome. METHODS: Data was downloaded from the TIMEx portal. TIMEx utilizes scRNA-seq signatures from 16 solid cancers to classify 37 cell types and activation states, including malignant, benign tissue, immune, and stromal cells. Unsupervised hierarchical clustering was performed to divide the TIMEx deconvoluted TCGA data (31 cancers, n=9848) into three clusters. Prevalence of each cancer type was calculated for each of the clusters. Progression free survival (PFS) was assessed by cluster for the entire cohort and for genitourinary (GU) cancers in a sub-analysis – Bladder (BLCA), Prostate (PRAD), Kidney [Clear Cell (KIRC), Papillary (KIRP), and Chromophobe (KICH)]. Tumor infiltrating lymphocyte (TIL) architecture data was collected from Saltz et al. (2018) in which “computational staining” was performed on the TCGA pathology slide archive. RESULTS: Three clusters with similar fractions of malignant and native tissue cells, but different immune and stromal enrichment were identified: CL1ImmuneLow/StromaLow, CL2ImmuneLow/StromaMod, and CL3ImmuneHi/StromaHi. While BLCA was heterogeneous, the majority of PRAD tumors were in CL1 (65%, p<2.2x10-16). The majority of KIRC tumors were in CL3 (80%, p<2.2x10-16), while KICH were mostly made up of CL1 (52%, p<2.2x10-16), and most KIRP were in CL2 (62%, p=2.2x10-13). Overall across cancers, CL3 had a longer PFS compared to grouped CL1 and CL2 combined (100.5 mo (95% CI 83.9-NR) vs. 85.7 mo (95% CI 74.4-101 mo), p=0.016). In a survival analysis of GU cancers, CL3 was associated with a longer PFS in BLCA (p=0.03) but shorter PFS in KIRC (p=0.05). Notably, BLCA CL3 patients had fewer FGFR mutations (p=0.007), brisk TIL spatial pattern (p=1.5x10-5), and more Basal/Squamous tumors by the Kamoun classifier (p=3.6x10-8) compared to CL1 and CL2. KIRC CL3 patients on the other hand, showed no difference in detection of VHL, PBRM1, SETD2, or TCEB1 alterations compared to CL1 and CL2. CONCLUSIONS: Our analyses reveal the heterogeneous composition and prognostic implications of the TME across various cancers. Further studies are needed to explore the capacity of this classification scheme to guide prognostication and therapeutic decisions. Source of Funding: A.M.U., S.S.S. are supported by the University of Michigan Health System-Peking University Health Science Center (UMHS-PUHSC) Joint Institute (JI). S.S.S. is supported by the Robert Wood Johnson Foundation as part of the Harold Amos Medical Faculty Development Program (AMFDP). S.S.S. is supported in part by the Urology Care Foundation Rising Stars in Urology Research Award Program and Astellas, Inc. S.S.S. is supported by P30CA046592. A.M.U. is supported by the Department of Defense (W81XWH-19-1-0407) and National Institutes of Health (U01 CA232931and R37 CA222829). A.M.U. and S.S.S. are supported by the National Institutes of Health (P50 CA186786) © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e34 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Mark Farha More articles by this author Brittney Cotta More articles by this author Srinivas Nallandhighal More articles by this author Kevin Hu More articles by this author Steven Goldenthal More articles by this author Aaron Udager More articles by this author Simpa Salami More articles by this author Expand All Advertisement PDF DownloadLoading ...

MP06-10 KDM6A DEFICIENCY IN UROTHELIUM PRODUCES A DISTINCT IMMUNE PHENOTYPE IN A MOUSE MODEL OF BLADDER CANCER

You have accessJournal of UrologyCME1 May 2022MP06-10 KDM6A DEFICIENCY IN UROTHELIUM PRODUCES A DISTINCT IMMUNE PHENOTYPE IN A MOUSE MODEL OF BLADDER CANCER Ao Zhang, Hong Qiu, Timothy Chan, and Byron Lee Ao ZhangAo Zhang More articles by this author , Hong QiuHong Qiu More articles by this author , Timothy ChanTimothy Chan More articles by this author , and Byron LeeByron Lee More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002523.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Lysine-specific demethylase 6A (KDM6A) is a chromatin modifier that is frequently mutated in bladder cancer and causes epigenetic dysregulation. Recently immune check point inhibition has shown great clinical promise in bladder cancer. Because epigenetic signature has been shown to predict response to immunotherapy, here we assessed if loss of KDM6A leads to alterations of tumor immune microenvironment in a mouse bladder cancer model. METHODS: Kdm6a urothelium conditional knockout (cKO) mice were generated by crossing Kdm6aflox/flox mice with Upk3aCre mice that express Cre recombinase in the urothelium. Littermate control mice were of the same genetic background but intact Kdm6a gene. Mice at 8 weeks of age (three in each group) were fed with drinking water containing 0.05% N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). Bladders were harvested after 4 weeks and analyzed with H&E staining. Single cell RNA sequencing was performed on urothelium from control and Kdm6A cKO mice without BBN treatment. RESULTS: After treatment with BBN for 4 weeks, there was marked edema and inflammatory changes with increased infiltration of immune cells in H&E stained bladder sections in Kdm6a cKO mice compared to Kdm6a intact controls (Figure A). While urothelium from control mice maintained grossly unchanged tissue architecture, urothelium from cKO mice exhibited cytologic atypia with loss of organized cell layer. Comparison of single cell RNA sequencing profile revealed enrichment of immune cell population in cKO mice (Figure B, cluster 12), which was consistent with histological findings. CONCLUSIONS: Loss of KDM6A in the urothelium creates a distinct immune microenvironment in the BBN mouse bladder cancer model. Whether this immune cell-rich microenvironment affects response to immune check point inhibition warrants further investigation. Source of Funding: This work was supported in part by the 2021 Urology Care Foundation Residency Research Award Program and the Kahlert Foundation © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e80 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ao Zhang More articles by this author Hong Qiu More articles by this author Timothy Chan More articles by this author Byron Lee More articles by this author Expand All Advertisement PDF DownloadLoading ...

Impact of a grant program to spur advances in sickle cell disease research

AbstractMore than 20 years ago, clinical trials and federal grant support for sickle cell disease (SCD) research were not on par with support for other genetic diseases. Faced with the opportunity to spur research and advance treatments for SCD, and at the recommendation of advisors, the Doris Duke Charitable Foundation (DDCF) offered an SCD research funding opportunity starting in 2009 through its Innovations in Clinical Research Awards (ICRA) program. Twenty-eight new grants of $450 000 for direct costs over 3 years and 7 renewals were awarded, for a total investment of $17 million. Only about half the research teams garnered follow-on funding directly related to their ICRA projects, but the financial return on the research investment was substantial (∼4 times the original $17 million or 300%). All but 1 of the ICRA investigative teams published original research reports that acknowledged DDCF as a source of funding; the median number of publications per team was 3. Major innovations in the diagnosis and treatment of SCD included but were not limited to a demonstration that genetic modification of BCL11A enhancer is a potentially important treatment modality, establishment that plerixafor mobilization is safe and effective for those with SCD, development and validation of a new diagnostic called SCD BioChip, and evidence that hydroxyurea treatment is safe and efficacious in African children. These outcomes show that relatively small research grants can have a substantial return on investment and result in significant advances for a disease such as SCD.

MP60-02 TRANSCRIPTOMICS CAN PREDICT HOMOLOGOUS RECOMBINATION DEFICIENCY IN PROSTATE CANCER

You have accessJournal of UrologyProstate Cancer: Markers (MP60)1 Sep 2021MP60-02 TRANSCRIPTOMICS CAN PREDICT HOMOLOGOUS RECOMBINATION DEFICIENCY IN PROSTATE CANCER Adam Weiner, Yang Liu, Matthew McFarlane, Pushpinder Bawa, Eric Li, Xin Zhao, Ziwen Li, Tanya Hammoud, Munna Hazime, R. Jeffrey Karnes, Elai Davicioni, Zachery Reichert, Arul Chinnaiyan, Tamara Lotan, and Edward Schaeffer Adam WeinerAdam Weiner More articles by this author , Yang LiuYang Liu More articles by this author , Matthew McFarlaneMatthew McFarlane More articles by this author , Pushpinder BawaPushpinder Bawa More articles by this author , Eric LiEric Li More articles by this author , Xin ZhaoXin Zhao More articles by this author , Ziwen LiZiwen Li More articles by this author , Tanya HammoudTanya Hammoud More articles by this author , Munna HazimeMunna Hazime More articles by this author , R. Jeffrey KarnesR. Jeffrey Karnes More articles by this author , Elai DavicioniElai Davicioni More articles by this author , Zachery ReichertZachery Reichert More articles by this author , Arul ChinnaiyanArul Chinnaiyan More articles by this author , Tamara LotanTamara Lotan More articles by this author , and Edward SchaefferEdward Schaeffer More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002095.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Tumors with mutations associated with homologous recombination deficiency (HRD) are uncommon in prostate cancer (PCa) and variably responsive to PARP inhibition. To better identify tumors with HRD, we developed and validated a transcriptomic signature for HRD in PCa (HRD-P). METHODS: By using an established mutational signature for tumor HRD, we created and validated a 16-gene signature for HRD-P in six independent PCa cohorts (primary PCa, n=8,224; metastatic castrate-resistant PCa [mCRPC], n=328). Molecular and clinical features were compared between HRD-P+ tumors and those with single HR-gene mutations that would have qualified patients for recent PARP inhibitor trials for mCRPC. RESULTS: HRD-P+ tumors were more numerous than tumors with single HR-gene mutations in primary (201/491, 41% vs 32/491 6.5%; Figure 1) and mCRPC (126/328, 38% vs 82/328, 25%) cases and HRD-P+ was more predictive of genomic instability suggestive of HRD. HRD-P+ was associated with shorter time to recurrence following surgery and shorter overall survival in men with mCRPC (Figure 2). In a prospective trial of mCRPC treated with olaparib (n=10), all three men with HRD-P+ experienced prolonged (>330 days) PSA progression-free survival. CONCLUSIONS: These results suggest transcriptomics can identify more patients that harbor phenotypic HRD than single HR-gene mutations and support further exploration of transcriptionally defined HRD tumors perhaps in conjunction with genomic markers for therapeutic application. Source of Funding: This work was supported in part by the National Institutes of Health grant 5U01CA196390 (EMS), the Prostate Cancer Foundation (EMS), Department of Defense grant W81XWH-15-1-0661 (EMS and TLL), the 2019 Urology Care Foundation Residency Research Award Program and the Russell Scott, Jr., MD Urology Research Fund (ABW), as well as the University of Michigan Prostate Specialized Program of Research Excellence (SPORE) P50 CA186786-05 and the Early Detection Research Network grant UO1 CA111275 (AMC). © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1041-e1041 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Adam Weiner More articles by this author Yang Liu More articles by this author Matthew McFarlane More articles by this author Pushpinder Bawa More articles by this author Eric Li More articles by this author Xin Zhao More articles by this author Ziwen Li More articles by this author Tanya Hammoud More articles by this author Munna Hazime More articles by this author R. Jeffrey Karnes More articles by this author Elai Davicioni More articles by this author Zachery Reichert More articles by this author Arul Chinnaiyan More articles by this author Tamara Lotan More articles by this author Edward Schaeffer More articles by this author Expand All Advertisement Loading ...

Impact of an institutional grant award on early career investigator applicants and peer reviewers

BackgroundObtaining research funding support is integral to a successful career in science. Training and practice in grant writing, as well as engagement in peer review of grant applications may help lead to successful research funding. However, there is little evidence on the impact of institutional programs on the career development of early career investigators (ECIs). ObjectivesUnderstand the impact of participation in an institutional research award program on the career development of ECIs. MethodsThe Cardiovascular Research Institute of Vermont established an Early Career Research (ECR) award program in 2018. ECIs who participated as applicants or reviewers in the first 3 years of the program (2018‐2020) were surveyed to understand the impact of the ECR award program on their grant writing and professional development. ResultsNinety‐four percent of 17 applicants and 90% of 19 reviewers completed the survey. Ninety‐two percent of funded and 75% of unfunded applicants, and 87% of reviewers reported that the program was beneficial to their professional development. Similarly, 85% of funded applicants, 75% of unfunded applicants, and 80% of reviewers reported improvement in their grant‐writing skills. All respondents reported they would recommend the ECR award program to their peers. ConclusionsThis single‐institution ECR award program had a positive impact on ECI’s professional development and grant‐writing skills and may lead to further extramural funding opportunities.

229-LB: Quantification of GLP-1R Internalization and Recycling in Live ß Cells

The glucagon-like peptide 1 receptor (GLP-1R) is a class B G-protein coupled receptor that is predominantly expressed on pancreatic β-cells in mice. Endogenous and synthetic ligands for GLP-1R exhibit distinct intracellular signaling cascades and promote insulin secretion to various degrees. Ligand binding typically induces receptor internalization followed by recycling back to the membrane or degradation within the lysosome. We hypothesized that endogenous and synthetic GLP-1R ligands would have distinct rates of internalization and recycling in β-cells. To test this hypothesis, we combined a recently-validated, highly-specific GLP-1R antibody (Glp1r0017) with flow cytometry and applied ligands (GLP-1, exendin-4 [Ex4], glucagon, and tirzepatide [TZP]) to characterize GLP-1R trafficking in freshly-isolated mouse islet cells. Under basal conditions, ~90% of β-cells were GLP1R+, with very low or absent binding in α- and δ-cells. Increasing ligand concentrations (0.01-100 nM) demonstrated distinct rates of internalization between ligands. Most potent was Ex4, followed by the native ligand GLP-1. TZP, a dual-receptor ligand for GLP-1R and GIPR, was more modest in inducing internalization. While showing different potencies, Ex4, GLP-1, and TZP were able to induce maximal internalization. In contrast, maximal treatment with glucagon only resulted in ~70% of β-cells being GLP-1R+. To test the rate of recycling back to the plasma membrane, we treated islet cells with Ex4 or GLP-1 with a dose that induced ~50% internalization, removed the ligand, and then monitored GLP1R+ β-cells over time. Within the first 15 minutes, GLP1R+ β-cells increased linearly and rapidly. This plateaued from 30-60 minutes, resulting in 60-70% of β-cells being GLP1R+ at the end of the experiment. In summary, our assay allows for investigation of GLP-1R trafficking in live β-cells. Understanding into the factors that regulate GLP-1R internalization and recycling may shed light on the insulinotropic properties of these ligands. Disclosure S. M. Gray: None. K. Sloop: Employee; Self; Eli Lilly and Company. P. Ravn: None. J. Campbell: None. D. A. D’alessio: Advisory Panel; Self; Eli Lilly and Company, Sun Pharmaceutical Industries Ltd., Research Support; Self; Eli Lilly and Company, Merck &amp; Co., Inc. Funding National Institutes of Health (F32DK121420, R01DK123075, R01DK101991); Lilly Research Award Program

A breath of fresh air - the potential for COVID-19 breath diagnostics.

A breath of fresh air - the potential for COVID-19 breath diagnostics.