Post-treatment Time Points Research Articles

A brief web-based depression literacy, efficacy, and stigma intervention among college students

ABSTRACT Objective The purpose of this study was to examine the impact of a brief web-based depression literacy intervention compared to a control condition on depression literacy, depression literacy efficacy, self-stigma, perceived public stigma, and personal stigma toward depression. Method College students (N = 210) from a large university in the southeastern United States were recruited via a human subject pool and randomly assigned to the intervention (web-based depression literacy) or control condition. Participants completed online surveys at pre-intervention, post-intervention, and 1-month follow-up time points. A short qualitative reflection question was also responded to post-intervention. Results No significant intervention condition by time interactions were seen across any variables included in the study. Significant main effects of time were found for the depression literacy, depression literacy efficacy, self-stigma, and personal stigma variables. Qualitative results show participants reported gaining a general understanding of depression in addition to more specific information such as knowing different treatment options or depression related resources. Discussion Education through a brief web-based depression literacy intervention did not improve college students’ literacy, confidence in recognising, managing, and preventing depression, or stigma towards this mental health disorder, compared to an active control condition.

Identification of optimal reference genes in golden Syrian hamster with ethanol- and palmitoleic acid-induced acute pancreatitis using quantitative real-time polymerase chain reaction.

Acute pancreatitis (AP) is a severe disorder that leads to high morbidity and mortality. Appropriate reference genes are important for gene analysis in AP. This study sought to study the expression stability of several reference genes in the golden Syrian hamster, a model of AP. AP was induced in golden Syrian hamster by intraperitoneal injection of ethanol (1.35 g/kg) and palmitoleic acid (2 mg/kg). The expression of candidate genes, including Actb, Gapdh, Eef2, Ywhaz, Rps18, Hprt1, Tubb, Rpl13a, Nono, and B2m, in hamster pancreas at different time points (1, 3, 6, 9, and 24 h) posttreatment was analyzed using quantitative polymerase chain reaction. The expression stability of these genes was calculated using BestKeeper, Comprehensive Delta CT, NormFinder, and geNorm algorithms and RefFinder software. Our results show that the expression of these reference genes fluctuated during AP, of which Ywhaz and Gapdh were the most stable genes, whereas Tubb, Eef2, and Actb were the least stable genes. Furthermore, these genes were used to normalize the expression of TNF-α messenger ribonucleic acid in inflamed pancreas. In conclusion, Ywhaz and Gapdh were suitable reference genes for gene expression analysis in AP induced in Syrian hamster.

A 2-year longitudinal randomized controlled trial examining the transfer of speed of processing training to secondary cognitive domains in middle-aged and older adults with HIV-associated neurocognitive disorder: Results of the think fast study

Objective: As people with HIV (PWH) age, they are at-risk of developing cognitive impairments compared to their seronegative counterparts. Although speed of processing (SOP) training may help improve this cognitive ability, less work has examined transfer to other cognitive domains. This study examined the effect of SOP training has on secondary cognitive domains in PWH aged 40+ years. Method: In this 3-group 2-year longitudinal study, 216 PWH with HIV-associated neurocognitive disorder (HAND) or borderline HAND were randomized to either: (1) 10 h of SOP training (n = 70); (2) 20 h of SOP training (n = 73); or (3) 10 h of an active control training (n = 73). Participants completed a comprehensive cognitive battery at baseline, immediately after training, and at 1 and 2 years. This battery yielded global and domain specific T-scores as well as a cognitive impairment variable. Generalized linear mixed-effect models were fitted to estimate between-group mean differences at the follow-up time-points adjusted for baseline. Results: No clinically or statistically significant improvements in any of the cognitive outcomes were observed. A sensitivity analysis was conducted; conclusions replicated those of the main analysis, with two exceptions: Global Function T and Psychomotor Speed T showed relevant training improvements among the intervention groups over the control group at the immediate post time point. Conclusions: Although SOP training has been shown to improve cognitive abilities that correspond to driving and mobility, such training has limited therapeutic utility in improving cognition in other domains in PWH with HAND.

Genetic events associated with venetoclax resistance in CLL identified by whole-exome sequencing of patient samples

AbstractAlthough BCL2 mutations are reported as later occurring events leading to venetoclax resistance, many other mechanisms of progression have been reported though remain poorly understood. Here, we analyze longitudinal tumor samples from 11 patients with disease progression while receiving venetoclax to characterize the clonal evolution of resistance. All patients tested showed increased in vitro resistance to venetoclax at the posttreatment time point. We found the previously described acquired BCL2-G101V mutation in only 4 of 11 patients, with 2 patients showing a very low variant allele fraction (0.03%-4.68%). Whole-exome sequencing revealed acquired loss(8p) in 4 of 11 patients, of which 2 patients also had gain (1q21.2-21.3) in the same cells affecting the MCL1 gene. In vitro experiments showed that CLL cells from the 4 patients with loss(8p) were more resistant to venetoclax than cells from those without it, with the cells from 2 patients also carrying gain (1q21.2-21.3) showing increased sensitivity to MCL1 inhibition. Progression samples with gain (1q21.2-21.3) were more susceptible to the combination of MCL1 inhibitor and venetoclax. Differential gene expression analysis comparing bulk RNA sequencing data from pretreatment and progression time points of all patients showed upregulation of proliferation, B-cell receptor (BCR), and NF-κB gene sets including MAPK genes. Cells from progression time points demonstrated upregulation of surface immunoglobulin M and higher pERK levels compared with those from the preprogression time point, suggesting an upregulation of BCR signaling that activates the MAPK pathway. Overall, our data suggest several mechanisms of acquired resistance to venetoclax in CLL that could pave the way for rationally designed combination treatments for patients with venetoclax-resistant CLL.

Cerebrospinal Fluid Biomarkers Associated with Intrathecal Cell Therapy in Patients with Progressive Multiple Sclerosis (P1-3.001)

<h3>Objective:</h3> To identify cerebrospinal fluid (CSF) biomarkers of clinical response to intrathecal (IT) mesenchymal stem cell-neural progenitor (MSC-NP) treatment in patients with progressive multiple sclerosis (MS). <h3>Background:</h3> MSC-NPs are a bone marrow-derived population of cells with trophic and immunomodulatory properties with therapeutic potential in MS. A phase II randomized, double-blind, placebo-controlled clinical trial was performed to investigate whether multiple intrathecal (IT) injections of autologous MSC-NPs was associated with clinical efficacy. Study subjects received either 6 IT injections of autologous MSC-NPs or 6 placebo injections spaced 2 months apart. In the crossover study design, subjects treated with placebo in the first year were treated with MSC-NPs in the second year, and vice versa. Safety and clinical efficacy will be reported elsewhere. <h3>Design/Methods:</h3> For biomarker discovery, CSF was collected from 36 study subjects at the time of their first and sixth IT-MSC-NP treatment reflecting pre-treatment and post-treatment timepoints, respectively. CSF supernatants were analyzed using SOMAScan multiplex proteomic assay. Differentially expressed proteins were identified using a multivariate linear mixed effect model. Biomarkers were validated using ELISA/Luminex assays. Treatment responders were defined by improvement in any of the following outcomes: EDSS, timed 25-foot walk, 9-hole peg test, 6-minute walk test, or muscle strength. <h3>Results:</h3> Proteomic analysis of CSF identified matrix metalloproteinase-9 (MMP-9), complement C3a and the chemokine CCL2 as potential biomarkers of IT-MSC-NP treatment. MMP-9 was significantly increased post-treatment and was observed independent of clinical response. Relative concentration of C3a, C3 and C3b were decreased post-treatment and subgroup analysis demonstrated significance only in the responder group. CCL2 was significantly decreased post-treatment with higher significance associated with clinical response. Decreased CCL2 was validated in an independent cohort of patients treated with IT-MSC-NPs. <h3>Conclusions:</h3> These results identify multiple CSF biomarkers of IT-MSC-NP treatment that may correlate with therapeutic response to this novel cell therapy in MS. <b>Disclosure:</b> Miss Kizilbash has nothing to disclose. Ms. Carlson has nothing to disclose. Violaine K. Harris, PhD has nothing to disclose. Dr. Sadiq has nothing to disclose.

The Development of a UK Culturally Adapted and Modified Version of the Person Attuned Musical Interactions Manual: Protocol for a 2-Phase Mixed Methods Study

Previous research has suggested that care home interactions need significant improvements, especially those between staff and residents with dementia. Reasons for the lack of interactions are staff time pressures and residents' language impairments. Although residents may experience reduced language abilities, they can continue to communicate through other forms, including nonverbal communication and music. Person Attuned Musical Interactions (PAMI) is a staff training tool that provides staff with music therapy skill-sharing to promote high-quality interactions between staff and residents using nonverbal communication and music. The tool was originally developed in Denmark. To ensure that the tool is appropriate for UK care homes, a team of researchers in the United Kingdom have modified and culturally adapted the tool. This study aims to investigate the appropriateness of the adapted and modified manual for UK care homes and to explore the impact of PAMI on residents with dementia and care staff. The project consists of 2 phases, a qualitative field-testing study and a mixed methods evaluation study, which have been developed following the Medical Research Council's guidelines for complex interventions. Care staff and residents with dementia will be recruited from care homes in Lincolnshire, where the care staff will be trained in the PAMI intervention before implementing the intervention in their daily routines. Fortnightly reflective sessions will be provided throughout the phases to provide supervision and monitoring. The qualitative methods include interviews, reflective session transcripts, diary entries, and resident experience questionnaires. The quantitative outcome measures are residents' music engagement, staff's dementia competence, residents' quality of life, and staff burden. The resident's music engagement will be administered at 9 fortnightly time points. Staff's dementia competence, resident's quality of life, and staff burden will be administered at preintervention and postintervention time points. The study has been funded by The Music Therapy Charity as part of a PhD studentship. The study began recruiting in September 2021. The research team aims to publish the results of the first phase in July to September 2023 and those of the second phase in October to December 2023. This study will be the first to investigate the modified version of PAMI. Therefore, it will provide feedback on the appropriateness of the manual for UK care homes. The PAMI intervention has the potential to offer high-quality music intervention training to a larger population of care homes who may currently be restricted by finances, the availability of time, and a lack of training opportunities. DERR1-10.2196/43408.

Effect of electroacupuncture treatment combined with rehabilitation care on serum sirt3 level and motor function in elderly patients with stroke hemiparesis.

Acupuncture treatment helps to improve neurological and motor function in elderly patients with stroke hemiplegia. However, the exact mechanism by which electroacupuncture improves stroke hemiparesis is uncertain. The aim of this study was to determine the effect of electroacupuncture care on sirt3 levels in elderly patients with stroke hemiparesis. One hundred and ten elderly patients with hemiplegia after first stroke were divided into an experimental group and a control group (n = 55 in each group). The control group was given conventional rehabilitation care by a rehabilitation therapist. In the experimental group, on the basis of conventional rehabilitation care, electroacupuncture was performed once a day for 28 days. Fugl-Meyer assessment (FMA) and barthel index (BI) scores were significantly higher, while neurologic deficit scale (NDS) and physiological state scores were significantly lower in both groups after 14 and 28 days of intervention compared to preintervention. The Generalized estimating equation (GEE) model also showed that the experimental group showed more favorable improvements in all outcomes at postintervention time points compared to the control group. After the intervention, serum sirt3 levels increased significantly in both groups compared to preintervention, and the increase was more pronounced in the experimental group. Consistently, the GEE model showed that serum sirt3 levels were significantly higher in the experimental group compared to the control group at postintervention time points. Correlation analysis revealed that serum sirt3 levels in the experimental group were negatively correlated with FMA and BI pre- and postintervention, while showing a significant positive correlation with NDS and physiological state scores. Electroacupuncture intervention led to significant improvements in motor function, activities of daily living and neurological function in elderly patients with stroke hemiplegia, which may be associate with increased serum sirt3 levels.

Abstract 2276: Spatiotemporal analysis of metastatic melanoma reveals mechanisms of resistance to immune checkpoint blockade

Abstract The treatment of metastatic melanoma has been revolutionized with the introduction of immune checkpoint blockade (ICB) therapies in the last decade. Despite durable responses exhibited by a subgroup of patients, most of patients do not respond to ICB therapies and the vast majority (60%-90%) of patients experience disease progression within five years. Molecular mechanisms of primary and acquired resistance to ICB are still widely unknown, which warrants further mechanistic investigation. Towards that goal, we used a holistic approach driven by integrated genomic tools to molecularly profile 35 longitudinal tumor specimens at pre-, on-, and post-treatment time points which were derived from seven patients with metastatic melanoma who progressed on sequential ICB therapies, including three responders and four non-responders. We integrated analyses of data generated from RNA sequencing, Whole-exome Sequencing (WES), NanoString nCounter vantage 3D, Cyclic Immunofluorescence (CyCIF), NanoString Digital Spatial Profiling (DSP) and Multiplex Immunohistochemistry (mIHC) platforms to elucidate evolutionary trajectories that occurred in both the tumor and the tumor immune microenvironment (TiME) that may have orchestrated response and resistance to ICB therapies. Simultaneous analyses conducted on genomic, transcriptomic and proteomic levels identified the re-activation of the MAPK pathway as a potential mechanism of resistance to ICB therapies. Spatially-resolved, image-based immune monitoring analysis at single cell resolution revealed that response to ICB is associated with infiltration of immune cells accompanied by induced activities of myeloid, T cells and macrophages in the TiME. In summary, integrated analyses allow us not only to reveal the dynamic co-revolution of both tumor and immune cells in TiME following sequential ICB therapies but also to provide a comprehensive perspective to molecular mechanisms of response and resistance to ICB therapies. A deeper understanding and elucidation of the evolution of mechanisms of response and resistance to ICB therapies will point us to generate hypothesis-driven, potential salvage therapies to overcome resistance to ICB therapies. Citation Format: Shiyou Wei, Jinho Lee, Marilyne Labrie, Courtney Betts, Lunxu Liu, Lisa Coussens, Meenhard Herlyn, Genevieve Boland, Gordon Mills, Gao Zhang. Spatiotemporal analysis of metastatic melanoma reveals mechanisms of resistance to immune checkpoint blockade [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2276.

Abstract 2282: Immune modulation of the T cell repertoire in mice following immunotherapy and chemotherapy combination treatments

Abstract Background: Immune checkpoint blockade therapeutics have revolutionized cancer therapy. However, most patients are either unresponsive or develop acquired resistance. Understanding patient-specific temporal T cell repertoires in the context of immune interventions will contribute to a better understanding of potential improvement strategies for better T cell anti-tumor immunity. Current methods to understand the T cell repertoire changes over time are limited due to the limited amount of material and prohibitive cost. In this study, we designed and performed longitudinal TCR sequencing in mice treated with multiple therapies. Methods: C57BL/6 mice were used in each treatment group, with blood drawn overtime before, during and after treatment. Mice were treated with a combination of immunotherapeutic agents and OVA peptide immunization, 100µl blood was drawn at multiple pre-, on- and post-treatment time points followed subsequently by TCR sequencing. Cyclophosphamide (CTX) is known to cause lymphodepletion followed by homeostatic proliferation. Thus, we also treated B16-F10 tumor bearing mice with CTX, and performed TCR sequencing on blood draws pre-, on-, and post-treatment to track TCR clone diversity and clonality over time. Results: We find that our method is sensitive enough to detect effects of immunotherapies on TCR diversity in small sample quantities. Using our method, we discovered that certain immunotherapeutic agents, most evidently CD40, have a significant but transient effect on the diversity of the T cell repertoire. We observed an initial increase in repertoire clonality, seven days post CD40 treatment followed by contraction. We show how we can identify antigen specific T cells and follow reacting T cell clones. In addition, we find that CTX treatment is associated with increased clonality in tumor bearing mice particularly in the week following treatment reflecting changes in the TCR repertoire. Conclusion: We have developed a method by which we can follow the TCR repertoire in mice treated with immunotherapy or chemotherapy. This enabled us to isolate and study the effects of different immune modulatory drugs of the endogenous T cell repertoire overtime. Citation Format: Yuval Elhanati, Mariam M. George, Hyejin Choi, Rachana Maniyar, Linda Hamadane, Neeman Mohibullah, Benjamin D. Greenbaum, Taha Merghoub. Immune modulation of the T cell repertoire in mice following immunotherapy and chemotherapy combination treatments [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2282.

Abstract 5588: Methylated ctDNA dynamics correspond with clinical tumor load in metastatic lung cancer patients on therapy

Abstract Treatment monitoring assays are needed to accurately and rapidly assess the efficacy of cancer treatments. While imaging remains the gold standard for monitoring the efficacy of cancer treatment, the use of more sensitive tools, such as liquid biopsy, could be beneficial for the patient’s ultimate treatment outcome. Several liquid biopsy-based assays that measure circulating tumor DNA (ctDNA) have been developed to meet this need. However, approaches that rely on quantifying the variant allele fraction (VAF) of somatic variants may be inaccurate or inconsistent due to a scarcity of detected somatic variants in the ctDNA or may be logistically infeasible if they require a tumor biopsy a priori. Methylated ctDNA has shown promise as a biomarker for treatment monitoring without requiring a tumor biopsy, but current efforts are limited in their ability to precisely quantify the amount of methylation present in the ctDNA. We hypothesize that more precise quantification of methylated ctDNA could enable more accurate correspondence with clinical tumor load and cancer treatment outcomes. Here, we present a retrospective study characterizing how amounts of methylated ctDNA dynamically change through cancer therapy. We tested 75 patient-treatment events, with each event composed of patient samples from one pre-treatment and two post-treatment time points, all from patients with Stage III-IV metastatic lung cancer receiving targeted therapies or immunotherapy. We employed a novel methylation-based, tumor-naive liquid biopsy assay for treatment monitoring by counting the number of methylated molecules in cell-free DNA. We observed methylation amounts ranging from &amp;lt;50 to &amp;gt;50,000 total methylated tumor molecules per 1000 assayed genomic equivalents at individual time points, indicating that the assay can detect a broad range of tumor load in blood. In addition, we measured changes between time points as large as &amp;gt;96% decrease and &amp;gt;2800% increase in methylation. We found that methylation correlated with maximum variant allele fractions as measured by a treatment selection assay (r2 = 0.74). Moreover, in this tumor-naive assay, we identified a median of 58 informative methylated loci, significantly more than a median of ~1-5 informative SNVs identified in VAF-based tumor-naive assays, and ~4-16 informative SNVs identified in VAF-based tumor-informed assays. This ~6-10x increase in the number of informative loci enables much more precise quantification and underlies the assay’s robustness and precision across patients, time points, and therapies. Finally, these changes in normalized total tumor molecules correlated with radiographic responses on imaging, duration of response, and time on therapy. Overall, our findings support the utility of methylated ctDNA as a biomarker for monitoring tumor load in cancer patients. Citation Format: Patrick Ye, Brian Woodward, Robb Viens, Sydne Langpap, Katherine Shelburne, Wen Zhou, Joyce Zhu, Jan Wignall, Gary Palmer, David Tsao, Oguzhan Atay, Hatim Husain. Methylated ctDNA dynamics correspond with clinical tumor load in metastatic lung cancer patients on therapy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5588.

Abstract 5247: The effect of aspirin on the transcriptional landscape of the colon

Abstract Colorectal cancer (CRC) remains the second leading cause of cancer mortality in the United States. Aspirin is the most promising CRC prevention agent with significant data supporting long-term use with a reduction in the incidence of adenomatous polyps and colorectal cancer. The adoption of aspirin as a preventive agent has been limited by an incomplete understanding of aspirin’s mode of action in humans. To elucidate the mechanism of aspirin prevention, we sought to identify aspirin-associated transcriptional changes within the colon. To accomplish this objective, we performed RNA sequencing of biopsy-derived colonic epithelial cells from participants enrolled (N=180) in our ongoing aspirin randomized placebo-control trial (RCT) program, “ASPirin Intervention for the REDuction of colorectal cancer risk” (ASPIRED). Individuals with a recent resection of an adenoma were eligible. Participants for whom epithelial cells from rectosigmoid mucosal biopsies were successfully obtained at both pre- and post-treatment timepoints (n=130) were included in this analysis. Participants were randomized to once-daily placebo (n=45), low-dose aspirin (81 mg) (n=46), or standard-dose aspirin (325 mg) (n=39) for 8-12 weeks. For each timepoint, we performed Illumina RNA 100-bp paired-end sequencing of 4,000 fluorescence-activated cell-sorted (FACS) colonic epithelial cells (CD45-; EpCAM+). Transcriptomic read alignment and read counts per gene were obtained using STAR and Salmon. Quality control measures removed 52 participants (20 placebo, 18 low-dose aspirin, and 14 standard-dose aspirin) from the analysis. No significant differences in age, gender, ethnicity, or BMI were observed between placebo and aspirin-treated participants. We then identified genes whose expression is modulated by aspirin exposure (low and standard dose aspirin) while controlling for baseline gene expression differences between placebo and aspirin-exposed patients and for intra-sample gene expression variability using Dream. We found 57 genes were significantly differentially expressed in aspirin-treated participants at an FDR &amp;lt; 0.10. AGR2, which is a CRC oncogene that is upregulated by prostaglandin E2 (PGE2), was significantly downregulated by aspirin treatment suggesting aspirin modules the expression of PGE2 responsive genes in the colonic epithelium. Gene enrichment analysis found aspirin significantly downregulated scaffold proteins (e.g. KRT8, KRT18, and ACTB) and structural ribosomal RNA binding proteins. These pathways are critical processes in epithelial cell differentiation. This finding supports previous in vitro evidence that aspirin results in cell composition changes within the colonic epithelium favoring stem-like cell states, which may underlie its preventive effect. Citation Format: Jonathan M. Downie, Connor M. Geraghty, Thaddeus S. Stappenbeck, Ömer H. Yilmaz, David A. Drew, Andrew T. Chan. The effect of aspirin on the transcriptional landscape of the colon. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5247.

45 Senolytic Therapy Transiently Reduces Inflammatory Markers in Primary Blood Mononuclear Cells of Individuals with Early Alzheimer’s Disease: Exploring the Conserved Transcriptional Response to Adversity as a Biomarker for Disease State

OBJECTIVES/GOALS: Determine if the Conserved Transcriptional Response to Adversity transcriptomic profile established in primary blood mononuclear cells (PBMC) of chronically stress caregivers, is present in individuals with early Alzheimer’s disease. Chronic stress is a risk factor for Alzheimer’s, and may be an untapped biomarker for disease risk and pathology. METHODS/STUDY POPULATION: To collect preliminary data on the Conserved Transcriptional Response to Adversity profile in individuals with Alzheimer’s disease, we were able to utilize primary blood mononuclear cell samples from a small open label pilot study called Senolytic Therapy to Modulate the Progression of Alzheimer’s Disease, designed to clear stressed senescent cells. We hypothesized senolytics may beneficially reverse this stress profile. We developed a NanoString assay (measuring 19 inflammatory, 31 type-1 interferon, and 3 antibody synthesis genes) to compare these transcriptomic changes within 4 individuals measured at baseline, post-treatment with an intermittent 12-week senolytic therapy, and at an optional extended post-treatment follow-up time point &gt; 3 months after their post treatment visit. RESULTS/ANTICIPATED RESULTS: There was relative downregulation of expression in transcription in 7 of 19 measured inflammatory genes (FOS, PTGS2, IL8, FOS, Il1b, JUNB, and JUN) in Alzheimer’s disease participants after receiving senolytic treatment (baseline vs. post-treatment). This is consistent with a decrease in the inflammatory arm of the Conserved Transcriptional Response to Adversity profile. These differences were not significant between baseline and the extended follow-up, indicative of a transient effect of senolytic. There were no changes in type 1 interferon or antibody synthesis genes. This data provides preliminary evidence for larger controlled studies to further establish this profile in Alzheimer’s disease, providing exciting evidence for transcript changes that may be reproducible with senolytic therapy. DISCUSSION/SIGNIFICANCE: Literature relevant to Alzheimer’s disease indicates global increases in inflammation paired with deficits in immune response, capturing some genes associated with the Conserved Transcriptional Response to Adversity. This profile may be a useful biomarker for prediction of disease severity or risk of dementia due to chronic stress.

Effects of Te Ara Mua – Future Streets suburban street retrofit on traffic speed and volume: Controlled before-after study

BackgroundTraffic speed and volume are important determinants of road traffic injury rates for all travel modes, and high traffic speed and volume can deter walking and cycling. Te Ara Mua – Future Streets is a project in Māngere, Aotearoa New Zealand, that aims to make it safer and easier for people to get around their local neighbourhood, especially by walking and cycling. Key components include implementation of a street hierarchy and street design changes to reduce speeds on neighbourhood streets, as well as other components including improved active transport infrastructure. MethodsWe used tube counters to measure motor vehicle speed and volume before and after the intervention, at different levels of the street hierarchy: arterial, collector and local streets. A difference-in-differences approach was used to assess changes in the intervention area compared with a matched control area. ResultsWe found post-intervention mean speed reductions of 8 km/h on local streets, and 5–6 km/h on collector streets, relative to the control area. Mean traffic volumes reduced by 17–24% on local streets, with no change on collectors or arterials. Both post-intervention time points showed similar results, indicating a sustained effect. ConclusionsOverall, this intervention successfully reinforced a street hierarchy, creating local residential streets that are more conducive to road safety and active travel. Implementation of similar interventions in disadvantaged neighbourhoods could help reduce road traffic injury inequities within cities. The findings suggest a promising contribution to safer streets that are more attractive for walking and cycling. Our future research will evaluate broader impacts of this intervention on travel mode shift, injury rates and other health and social outcomes.

Assessment of a novel continuous cleaning device using metatranscriptomics in diverse hospital environments.

Despite routine implementation of cleaning and disinfection practices in clinical healthcare settings, high-touch environmental surfaces and contaminated equipment often serve as reservoirs for the transmission of pathogens associated with healthcare-associated infections (HAIs). The current study involved the analysis of high-touch surface swabs using a metatranscriptomic sequencing workflow (CSI-Dx™) to assess the efficacy of cleanSURFACES® technology in decreasing microbial burden by limiting re-contamination. This is a non-human single center study conducted in the Emergency Department (ED) and on an inpatient Oncology Ward of Walter Reed National Military Medical Center that have followed hygienic practices during the COVID-19 pandemic environment. Although there was no difference in observed microbial richness (two-tailed Wilcoxon test with Holm correction, P > 0.05), beta diversity findings identified shifts in microbial community structure between surfaces from baseline and post-intervention timepoints (Day 1, Day 7, Day 14, and Day 28). Biomarker and regression analyses identified significant reductions in annotated transcripts for various clinically relevant microorganisms' post-intervention, coagulase-negative staphylococci and Malassezia restricta, at ED and Oncology ward, respectively. Additionally, post-intervention samples predominantly consisted of Proteobacteria and to a lesser extent skin commensals and endogenous environmental microorganisms in both departments. Findings support the value of cleanSURFACES®, when coupled with routine disinfection practices, to effectively impact on the composition of active microbial communities found on high-touch surfaces in two different patient care areas of the hospital (one outpatient and one inpatient) with unique demands and patient-centered practices.

Abstract P2-21-02: Longitudinal tumor-immune microenvironment changes in patients with clinical luminal A early breast cancer treated with neoadjuvant palbociclib and endocrine therapy: results from the Swedish randomized PREDIX Luminal A trial

Abstract Introduction: The use of preoperative CDK4/6 inhibitors combined with endocrine treatment remains investigational in breast cancer (BC), while their effect on tumor-immune microenvironment (TIME) in luminal BC is the subject of few published studies. The aim of this study was to characterize the treatment-induced changes of TIME at a spatial proteomic and bulk tumor transcriptomic level in clinical luminal A BC patients treated with neoadjuvant palbociclib and endocrine treatment. Methods: The PREDIX Luminal A phase II randomized clinical trial (NCT02592083) included patients with stage 2-3 BC and IHC-defined luminal A biology (ER+ and PR+, HER2-, low proliferation) treated with preoperative ET (tamoxifen or aromatase inhibitor) with or without the addition of CDK4/6i palbociclib, based on the early change of Ki67 levels. A core biopsy was obtained at baseline, after 10 weeks of treatment (on-treatment) and at surgery. The study was closed after the inclusion of 10 patients due to slow accrual. By using formalin-fixed paraffin-embedded tissue from all timepoints, we performed the GeoMx® Digital Spatial Profiling (DSP, NanoString, Seattle, WA, USA) for the spatial profiling of 27 immune cell and tumor markers (immune cell profiling core and pan-tumor panels) both at intra-epithelial and stromal tissue segments. Upon data normalization, a linear mixed model was adopted for differential marker expression assessment between the different timepoints and treatment arms, using the DSP Suite® software. RNA was also extracted from fresh-frozen biopsies and the nCounter® Breast Cancer 360™ panel (NanoString) was used for gene expression profiling of 776 targets and data analysis was performed using the nSolver® software. Results: A total of 7 patients received palbociclib + ET and 3 were treated with ET only. All patients experienced an on-treatment radiological partial response while no patient achieved pathological complete response. While ET suppressed the ER axis in both palbociclib-treated and untreated patients, treatment with palbociclib additionally led to cell-cycle arrest as evidenced by a decrease in proliferation and downregulation of cell-cycle gene pathways. Immune-related protein markers were enriched in the stroma segments and pan-tumor markers were enriched in the intra-epithelial segments at all timepoints. In palbociclib-treated patients, immune cell markers (e.g. CD45, CD68, CD20, CD4, HLA-DR) were significantly enriched in on-treatment samples (n=3) both in tumor and stromal segments, while these expression changes were reversed between on-treatment and surgery. In the ET-treated patients immune markers reflecting e.g. T-, B- and/or antigen-presenting-cells were significantly enriched at the operation samples (n=2), in the intra-epithelial but not in the stromal areas. This finding was confirmed also at the gene level, where immune-related pathways (e.g. chemokine and cytokine activity and receptor binding) were upregulated in the palbociclib-treated patients on-treatment but they were partially abrogated at surgery. The ET-only treated patients presented with enhanced immune activation gene sets at both on- and post-treatment timepoints Conclusion: The findings of this small hypothesis-generating study indicate an upregulation of immune function during treatment with palbociclib and ET and an altered tumor microenvironment. The prognostic and potential therapeutic implications of sensitizing tumors to subsequent chemotherapy and immunotherapy need to be further investigated in larger studies. Citation Format: Ioannis Zerdes, Dimitrios Salgkamis, Alexios Matikas, Lars Selander, Kang Wang, Evangelos Tzoras, Emmanouil Sifakis, Susanne Agartz, Xinsong Chen, Mats Hellström, Johan Hartman, Jonas Bergh, Theodoros Foukakis, Thomas Hatschek. Longitudinal tumor-immune microenvironment changes in patients with clinical luminal A early breast cancer treated with neoadjuvant palbociclib and endocrine therapy: results from the Swedish randomized PREDIX Luminal A trial [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-21-02.

Management of severe scoliosis in patients with Turner's syndrome: A case series

Aims and objectivesThe prevalence and treatment of severe scoliosis and other spinal anomalies in patients with Turner's syndrome (TS) is not well reported. This is the largest case series to date regarding the treatment course and outcomes of severely scoliotic TS patients. MethodsA retrospective chart review was performed to identify all patients with TS seen at a single center academic pediatric institution from 2007 to 2021. Of these, the presence of concomitant severe scoliosis or other spinal anomalies was determined, defined by a major coronal curve measuring 45° or greater. Demographic, clinical, surgical, and radiologic data was collected at both pre- and post-intervention time points. ResultsA retrospective chart review identified 306 patients with TS. Of those, six were identified to have severe scoliosis or other severe spinal anomalies requiring fusion. All four posterior spinal fusion (PSF) patients demonstrated improvement of their spinal curvature. One patient who electively pursued only bracing demonstrated minimal improvement and surgery was subsequently recommended, but not pursued. One patient expired from a pre-existing heart condition prior to intervention. All postoperative complications resolved with no further complications. The only brace-related complication was an allergic rash related to the brace material. ConclusionAll four patients who underwent PSF demonstrated significant improvement of their spinal curvature with few post-surgical complications. None of the patients in the bracing cohort demonstrated stabilization of their spinal curvature. Therefore, these data corroborate with prior studies, suggesting that operative management consisting of spinal fusion with instrumentation provides optimal clinical outcomes, compared to bracing only.

Metabolomic identification of predictive and early biomarkers of cisplatin-induced acute kidney injury in adult head and neck cancer patients.

Cisplatin causes acute kidney injury (AKI) in approximately one third of patients. Serum creatinine and urinary output are poor markers of cisplatin-induced AKI. Metabolomics was utilized to identify predictive or early diagnostic biomarkers of cisplatin-induced AKI. Thirty-one adult head and neck cancer patients receiving cisplatin (dose ≥70 mg/m2) were recruited for metabolomics analysis. Urine and serum samples were collected prior to cisplatin (pre), 24-48 h after cisplatin (24-48 h) and 5-14 days (post) after cisplatin. Based on serum creatinine concentrations measured at the post timepoint, 11/31 patients were classified with clinical AKI. Untargeted metabolomics was performed using liquid chromatography-mass spectrometry (LC-MS). Metabolic discrimination was observed between "AKI" patients and "no AKI" patients at all timepoints. Urinary glycine, hippuric acid sulfate, 3-hydroxydecanedioc acid and suberate were significantly different between AKI patients and no AKI patients prior to cisplatin infusion. Urinary glycine and hippuric acid sulfate were lower (-2.22-fold and -8.85-fold), whereas 3-hydroxydecanedioc acid and suberate were higher (3.62-fold and 1.91-fold) in AKI patients relative to no AKI patients. Several urine and serum metabolites were found to be altered 24-48 h following cisplatin infusion, particularly metabolites involved with mitochondrial energetics. We propose glycine, hippuric acid sulfate, 3-hydroxydecanedioc acid and suberate as predictive biomarkers of predisposition to cisplatin-induced AKI. Metabolites indicative of mitochondrial dysfunction may serve as early markers of subclinical AKI.

Oxylipin status, before and after LC n-3 PUFA supplementation, has little relationship with skeletal muscle biology in older adults at risk of sarcopenia

IntroductionOxylipins form endogenously via the oxygenation of long-chain polyunsaturated fatty acids (LC PUFA). Several oxylipins are highly bioactive molecules and are believed to be key mediators of LC PUFA metabolism in the body. However, little is known in relation to whether oxylipins mediate alterations in skeletal muscle mass and function. The objective of this study was to determine if a relationship exists between the oxylipin profile and skeletal muscle biology in healthy older adults at risk of sarcopenia and determine if this changes in response to LC n-3 PUFA supplementation. Materials and methodsThis exploratory study investigated the baseline correlations between LC n-3, n-6 and n-9 PUFA-derived oxylipins and markers of muscle biology. For this, the concentration of 79 free (i.e., non-esterified) oxylipins was quantified in human plasma by liquid chromatography-mass spectrometry (LC-MS) and retrospectively correlated to phenotypic outcomes obtained pre-intervention from the NUTRIMAL study (n = 49). After examining the baseline relationship, the potential effect of supplementation (LC n-3 PUFA or an isoenergetic control made of high-oleic sunflower and corn oil) was evaluated by correlating the change in oxylipins concentration and the change in markers of skeletal muscle biology. The relationship between oxylipins pre- and post-intervention and their parent PUFA were also examined. ResultsAt baseline, the hydroxy product of mead acid (n-9 PUFA), 5-HETrE, was negatively correlated to the phenotypic parameters appendicular lean mass index (ALMI) (p = 0.003, r=-0.41), skeletal muscle mass index (SMMI) (p = 0.001, r=-0.46), handgrip strength (HGS) (p<0.001, r = 0.48) and isometric knee extension (p<0.001, r=-0.48). Likewise, LC n-6 PUFA hydroxy‑PUFA were negatively correlated to HGS (i.e., 12-HETrE, p = 0.002, r=-0.42, and 5- and 11-HETE, p = 0.006, r=-0.47 and p<0.001, r=-0.50 respectively), single leg stand time (i.e., 12-HETrE, p = 0.006, r=-0.39 and 16-HETE, p = 0.002, r=-0.43), and five-time-sit-to-stand test (FTST) performance (16-HETE, p = 0.006, r = 0.39), and positively correlated to gait speed (i.e., 12-HETrE, p = 0.007, r = 0.38 and 16-HETE, p = 0.006, r = 0.39). LC n-3 PUFA supplementation increased eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) derived oxylipins and reduced n-6 PUFA derived oxylipins. Parameters of skeletal muscle mass and strength were not significantly altered in either LC n-3 PUFA or placebo groups. Changes in plasma oxylipins concentrations were closely related to changes in their parent PUFA, assessed in the erythrocyte membrane, but were not associated with any changes in skeletal muscle parameters. Discussion and conclusionAt baseline, the status n-9 (5-HETrE) and n-6 PUFA derivates [12-HETrE, and 5-, 11- and 16-HETE], but not n-3 PUFA derived oxylipins, were associated with poor skeletal muscle health parameters (i.e., mass and strength). However, these correlations were no longer present when correlating relative changes from pre to post timepoints. An independent cohort validation is needed to explore baseline correlations further. Further research is warranted to assess other biological mechanisms by which LC n-3 PUFA might affect muscle biology.

Healing From Home: Examination of an Online Mindfulness-Based Tinnitus Stress Reduction Course During the 2020 COVID Pandemic.

The purpose of this study was to assess the effectiveness of an Internet delivered Mindfulness Based Tinnitus Stress Reduction (i-MBTSR) program during the COVID-19 pandemic. Tinnitus, often a chronic condition, is experienced by 15% of the population. For those with severe, bothersome tinnitus, quality of life is reduced and the treatment options are limited. We evaluated an approach intended to decrease tinnitus-related distress. This study examined the data collected for an online i-MBTSR course using a retrospective design. The intervention included an 8-week self-paced i-MBTSR course, including didactic information about mindfulness and tinnitus, as well as meditation practices. Outcome measures included the Tinnitus Functional Index (TFI) and Perceived Stress Scale (PSS), which were measured at four time periods. These time points included pretreatment, midtreatment at 3 weeks, immediately posttreatment, and at 6-month follow-up. Forty-three participants completed the intervention. The mean preintervention tinnitus severity rating was 59.96 ("severe tinnitus") as measured by the TFI. Mean TFI scores dropped to 44.16 (p < .001) at midtreatment and to 34.23 (p = .001) at posttreatment. Repeated-measures analysis of variance and multivariate analysis of variance tests were conducted to determine changes in the two scales at the four time periods. There were significant differences in perceived stress and tinnitus self-function found in all measures and submeasures between the pre-, mid-, and posttreatment time points with the exception of the TFI Auditory subtest. These gains remained significant for those who completed the 6-month follow-up. The i-MBTSR course appears to be a viable and effective treatment modality. A shorter 3-week course may be effective. Case-control studies to more systematically investigate the effectiveness of i-MBTSR for tinnitus are required.

A Prospective, Open-Label Study to Evaluate Functional Changes Following Electromagnetic Muscle Stimulation of Abdominal Muscle.

Electromagnetic muscle stimulation (EMMS) is an effective, well-tolerated noninvasive body contouring treatment for strengthening, toning, and firming the abdomen. In this study, functional changes following abdominal EMMS treatment wereevaluated. In this prospective, open-label study, adults received 8 abdominal EMMS treatments (2 treatments on nonconsecutive days/week over 4 weeks). Follow-ups occurred 1 month (primary endpoint), 2, and 3 months postfinal treatment. Effectiveness endpoints included improvements from baseline on Body Satisfaction Questionnaire (BSQ; primary endpoint), core strength (timed plank test), abdominal endurance (curl-up test), and Subject Experience Questionnaire (SEQ). Safety was evaluated throughout. Sixteen participants (68.8% female) were enrolled, with a mean age of 39.3 years and a mean BMI of 24.4 kg/m2; 14 participants completed the study per protocol. Mean BSQ scores were significantly improved from baseline (27.9) to the 1-month follow-up (36.6; P < .05). Core strength and abdominal endurance were significantly greater at the 1-, 2-, and 3-month posttreatment time points than at baseline (P < .05). Frequently cited reasons for seeking EMMS treatment included a desire to feel stronger (100%; n = 14/14) and to improve athletic performance (100%; n = 14/14). SEQ responses 3 months posttreatment showed that most participants reported feeling stronger (92.9%) and motivated to receive additional EMMS treatments (100%) and work out to maintain treatment results (100%). The majority of participants (>78%) reported being "satisfied" or "very satisfied" with abdominal treatment 1 month posttreatment. One device- and/or procedure-related adverse event of menstrual cycle irregularity was reported in 1 participant and was mild in severity. EMMS treatment of the abdomen is associated with functional strength improvements and high patient satisfaction.