Changes In Immune Function Research Articles

Inflammatory gene expression during acute high‐altitude exposure

The molecular signalling pathways that regulate inflammation and the response to hypoxia share significant crosstalk and appear to play major roles in high‐altitude acclimatization and adaptation. Several studies demonstrate increases in circulating candidate inflammatory markers during acute high‐altitude exposure, but significant gaps remain in our understanding of how inflammation and immune function change at high altitude and whether these responses contribute to high‐altitude pathologies, such as acute mountain sickness. To address this, we took an unbiased transcriptomic approach, including RNA sequencing and direct digital mRNA detection with NanoString, to identify changes in the inflammatory profile of peripheral blood throughout 3 days of high‐altitude acclimatization in healthy sea‐level residents (n = 15; five women). Several inflammation‐related genes were upregulated on the first day of high‐altitude exposure, including a large increase in HMGB1 (high mobility group box 1), a damage‐associated molecular pattern (DAMP) molecule that amplifies immune responses during tissue injury. Differentially expressed genes on the first and third days of acclimatization were enriched for several inflammatory pathways, including nuclear factor‐κB and Toll‐like receptor (TLR) signalling. Indeed, both TLR4 and LY96, which encodes the lipopolysaccharide binding protein (MD‐2), were upregulated at high altitude. Finally, FASLG and SMAD7 were associated with acute mountain sickness scores and peripheral oxygen saturation levels on the first day at high altitude, suggesting a potential role of immune regulation in response to high‐altitude hypoxia. These results indicate that acute high‐altitude exposure upregulates inflammatory signalling pathways and might sensitize the TLR4 signalling pathway to subsequent inflammatory stimuli. Key points Inflammation plays a crucial role in the physiological response to hypoxia.High‐altitude hypoxia exposure causes alterations in the inflammatory profile that might play an adaptive or maladaptive role in acclimatization.In this study, we characterized changes in the inflammatory profile following acute high‐altitude exposure.We report upregulation of novel inflammation‐related genes in the first 3 days of high‐altitude exposure, which might play a role in immune system sensitization.These results provide insight into how hypoxia‐induced inflammation might contribute to high‐altitude pathologies and exacerbate inflammatory responses in critical illnesses associated with hypoxaemia.

A Novel Method for Identifying the Transition Zone in Long-Segment Hirschsprung Disease: Investigating the Muscle Unit to Ganglion Ratio.

Hirschsprung disease (HSCR) is characterised by the absence of enteric ganglia along variable lengths of the distal bowel. Current gold standard treatment involves the surgical resection of the defective, aganglionic bowel. Clear and reliable distinction of the normoganglionated bowel from the transition zone is key for successful resection of the entire defective bowel, and the avoidance of subsequent postoperative complications. However, the intraoperative nature of the tissue analysis and the variability of patient samples, sample preparation, and operator objectivity, make reproducible identification of the transition zone difficult. Here, we have described a novel method for using muscle units as a distinctive landmark for quantifying the density of enteric ganglia in resection specimens from HSCR patients. We show that the muscle unit to ganglion ratio is greater in the transition zone when compared with the proximal, normoganglionated region for long-segment HSCR patients. Patients with short-segment HSCR were also investigated, however, the muscle unit to ganglion ratio was not significantly different in these patients. Immunohistochemical examination of individual ganglia showed that there were no differences in the proportions of either enteric neurons or glial cells through the different regions of the resected colon. In addition, we identified that the size of enteric ganglia was smaller for patients that went on to develop HSCR associated enterocolitis; although the density of ganglia, as determined by the muscle unit to ganglia ratio, was not different when compared with patients that had no further complications. This suggests that subtle changes in the enteric nervous system, even in the “normoganglionated” colon, could be involved in changes in immune function and subsequent bacterial dysbiosis.

Distinct immune-effector and metabolic profile of CD8+ T cells in patients with autoimmune polyarthritis induced by therapy with immune checkpoint inhibitors

ObjectivesRheumatic immune-related adverse events (irAE) such as (poly)arthritis in patients undergoing immune checkpoint inhibitor (ICI) treatment pose a major clinical challenge. ICI therapy improves CD8+ T cell (CD8) function, but...

A Novel Purine and Uric Metabolism Signature Predicting the Prognosis of Hepatocellular Carcinoma.

Background: Hepatocellular carcinoma (HCC) is regarded as one of the most common cancers in the world with a poor prognosis. Patients with HCC often have abnormal purine and uric acid metabolism, but their relationship with prognosis is unclear. Methods: Here, we collected the data of peripheral blood uric acid and clinical data in 50 patients with HCC and analyzed the relationship with prognosis. At the same time, the transcriptome sequencing data of TCGA and GEO databases were collected to analyze the changes in purine metabolic pathway activity and construct a prognosis prediction model. Based on the prognosis prediction model related to purine metabolism, we further looked for the differences in the immune microenvironment and molecular level and provided possible drug targets. Results: We found that the level of serum uric acid was positively correlated with the prognosis of HCC. At the same time, purine metabolism and purine biosynthesis pathway activities were significantly activated in patients with a poor prognosis of HCC. The prognosis prediction model of HCC based on purine metabolism and purine biosynthesis pathway can accurately evaluate the prognosis of patients with HCC. Meanwhile, we found that there were significant changes in tumor immune infiltration microenvironment and biological function at the molecular level in patients with over-activation of purine metabolism and purine biosynthesis pathway. In addition, we found that uric acid level was positively correlated with peripheral blood leukocytes in HCC patients. Conclusion: In this study, we found that the level of peripheral blood uric acid in patients with HCC is correlated with their prognosis. The prognosis of patients with HCC can be accurately predicted through the metabolic process of uric acid and purine.

Sphingosine Phosphate Lyase Is Upregulated in Duchenne Muscular Dystrophy, and Its Inhibition Early in Life Attenuates Inflammation and Dystrophy in Mdx Mice.

Duchenne muscular dystrophy (DMD) is a congenital myopathy caused by mutations in the dystrophin gene. DMD pathology is marked by myositis, muscle fiber degeneration, and eventual muscle replacement by fibrosis and adipose tissue. Satellite cells (SC) are muscle stem cells critical for muscle regeneration. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid that promotes SC proliferation, regulates lymphocyte trafficking, and is irreversibly degraded by sphingosine phosphate lyase (SPL). Here, we show that SPL is virtually absent in normal human and murine skeletal muscle but highly expressed in inflammatory infiltrates and degenerating fibers of dystrophic DMD muscle. In mdx mice that model DMD, high SPL expression is correlated with dysregulated S1P metabolism. Perinatal delivery of the SPL inhibitor LX2931 to mdx mice augmented muscle S1P and SC numbers, reduced leukocytes in peripheral blood and skeletal muscle, and attenuated muscle inflammation and degeneration. The effect on SC was also observed in SCID/mdx mice that lack mature T and B lymphocytes. Transcriptional profiling in the skeletal muscles of LX2931-treated vs. control mdx mice demonstrated changes in innate and adaptive immune functions, plasma membrane interactions with the extracellular matrix (ECM), and axon guidance, a known function of SC. Our cumulative findings suggest that by raising muscle S1P and simultaneously disrupting the chemotactic gradient required for lymphocyte egress, SPL inhibition exerts a combination of muscle-intrinsic and systemic effects that are beneficial in the context of muscular dystrophy.

Psychological Stress Perceived by Pregnant Women in the Last Trimester of Pregnancy.

Pregnancy is characterized by changes in neuroendocrine, cardiovascular, and immune function. For this reason, pregnancy itself is perceived as a psychological “stress test”. Research to date has focused on stress exposure. The aim of the study was to evaluate the influence of associated factors on the level of stress experienced by pregnant patients. We conducted a prospective study that included 215 pregnant women in the third trimester of pregnancy, hospitalized in the Obstetrics and Gynecology Clinic II in Târgu-Mureș, between December 2019 and December 2021, who were evaluated by the ABS II scale. All patients included in the study filled in a questionnaire that included 76 questions/items, in which all the data necessary for the study were recorded. The results obtained from the study showed that pregnant women in urban areas (53.49%) are more vulnerable than those in rural areas (46.51%), being influenced by social and professional stressors, social determinants playing a critical role in pregnancy and in the newborn. Patients who have had an imminent abortion in their current pregnancy have a significantly higher score of irrationality than those with normal pregnancy, which shows that their emotional state can negatively influence the phenomenon of irrationality. There is a statistically significant association between pregnancy type I (normal pregnancy or imminent pregnancy) and irrationality class (p = 0.0001; RR: 2.150, CI (95%): 1.154–4.007). In the case of women with desired pregnancies, the risk of developing irrationality class IV–V is 4.739 times higher, with the association being statistically significant (p < 0.0001; RR 4.739; CI (95%): 2.144–10.476). The analysis of the obtained results demonstrates the importance of contributing factors and identifies the possibility of stress disorders, occurring in the last trimester of pregnancy, disorders that can have direct effects on maternal and fetal health. We consider it extremely important to carry out evaluations throughout the pregnancy. At the same time, it is necessary to introduce a screening program to provide psychological counseling in the prenatal care of expectant mothers.

The Blood Immune Cell Count, Immunoglobulin, Inflammatory Factor, and Milk Trace Element in Transition Cows and Calves Were Altered by Increasing the Dietary n-3 or n-6 Polyunsaturated Fatty Acid Levels.

Transition dairy cows experience sudden changes in both metabolic and immune functions, which lead to many diseases in postpartum cows. Therefore, it is crucial to monitor and guarantee the nutritional and healthy status of transition cows. The objective of this study was to determine the effect of diet enriched in n-3 or n-6 polyunsaturated fatty acid (PUFA) on colostrum composition and blood immune index of multiparous Holstein cows and neonatal calves during the transition period. Forty-five multiparous Holstein dairy cows at 240 days of pregnancy were randomly assigned to receive 1 of 3 isoenergetic and isoprotein diets: 1) CON, hydrogenated fatty acid (control), 1% of hydrogenated fatty acid [diet dry matter (DM) basis] during prepartum and postpartum, respectively; 2) HN3, 3.5% of extruding flaxseed (diet DM basis, n-3 PUFA source); 3) HN6, 8% of extruding soybeans (diet DM basis, C18:2n-6 PUFA source). Diets containing n-3 and n-6 PUFA sources decreased colostrum immunoglobulin G (IgG) concentration but did not significantly change the colostrum IgG yield compared with those with CON. The commercial milk yield (from 14 to 28 days after calving) was higher in the HN3 and HN6 than that in the CON. Furthermore, the n-3 PUFA source increased neutrophil cell counts in blood during the prepartum period and increased neutrophil percentage during the postpartum period when compared with those with control treatment. Diets containing supplemental n-3 PUFA decreased the serum concentration of interleukin (IL)-1β in maternal cows compared with those in control and n-6 PUFA during prepartum and postpartum. In addition, the neonatal calf serum concentration of tumor necrosis factor (TNF) was decreased in HN3 compared with that in the HN6 treatment. The diet with the n-3 PUFA source could potentially increase the capacity of neutrophils to defend against pathogens in maternal cows by increasing the neutrophil numbers and percentage during the transition period. Meanwhile, the diet with n-3 PUFA source could decrease the pro-inflammatary cytokine IL-1β of maternal cows during the transition period and decline the content of pro-inflammatary cytokine TNF of neonatal calves. It suggested that the highest milk production in n-3 PUFA treatment may partially be due to these beneficial alterations.

Immunosenescence of brain accelerates Alzheimer's disease progression.

Most of Alzheimer's disease (AD) cases are sporadic and occur after age 65. With prolonged life expectancy and general population aging, AD is becoming a significant public health concern. The immune system supports brain development, plasticity, and homeostasis, yet it is particularly vulnerable to aging-related changes. Aging of the immune system, called immunosenescence, is the multifaceted remodeling of the immune system during aging. Immunosenescence is a contributing factor to various age-related diseases, including AD. Age-related changes in brain immune cell phenotype and function, crosstalk between immune cells and neural cells, and neuroinflammation work together to promote neurodegeneration and age-related cognitive impairment. Although numerous studies have confirmed the correlation between systemic immune changes and AD, few studies focus on the immune state of brain microenvironment in aging and AD. This review mainly addresses the changes of brain immune microenvironment in aging and AD. Specifically, we delineate how various aspects of the brain immune microenvironment, including immune gateways, immune cells, and molecules, and the interplay between immune cells and neural cells, accelerate AD pathogenesis during aging. We also propose a theoretical framework of therapeutic strategies selectively targeting the different mechanisms to restore brain immune homeostasis.

Immune Checkpoint Inhibitors: The Unexplored Landscape of Geriatric Oncology.

Cancer is classically considered a disease of aging, with over half of all new cancer diagnoses occurring in patients over the age of 65 years. Immune checkpoint inhibitors (ICIs) have revolutionized cancer treatment, yet the participation of older adults with cancer in ICI trials has been suboptimal, particularly at the extremes of age. Despite significant improvement in treatment response and an improved toxicity profile when compared with conventional cytotoxic chemotherapies, many cancers develop resistance to ICIs, and these drugs are not free of toxicities. This becomes particularly important in the setting of older adults with cancer, who are generally frailer and harbor more comorbidities than do their younger counterparts. Immunosenescence, a concept involving age-related changes in immune function, may also play a role in differential responses to ICI treatment in older patients. Data on ICI treatment response in older adult with cancers remains inconclusive, with multiple studies revealing conflicting results. The molecular mechanisms underlying response to ICIs in older cancer patients are poorly understood, and predictors of response that can delineate responders from non-responders remain to be elucidated. In this review, we explore the unique geriatric oncology population by analyzing existing retrospective datasets, and we also sought to highlight potential cellular, inflammatory, and molecular changes associated with aging as potential biomarkers for response to ICIs.

Enhanced Tumoricidal Immune Responses by Transarterial Chemotherapy Using Novel Nanocomplexes in a Rat Liver Cancer Model.

Locoregional treatments for hepatocellular carcinoma (HCC) induce immunogenic cell death and a tumor-specific immune response, but infiltration and activation of immune cells in the liver have not been clearly described. Transarterial chemoembolization (TACE) or transarterial chemotherapy (TAC) without embolization have been used to treat intermediate or advanced stage HCC patients. The identification of intrahepatic immune cell changes after locoregional therapy provides a theoretical basis for the combination with immune checkpoint inhibitors (ICIs) in HCC. This study aimed to determine the anticancer effect and changes in the liver immune cell population and function after direct injection of polymerized phenylboronic acid-conjugated doxorubicin (pPBA-Dox) nanocomplexes into the liver through TAC. pPBA-Dox nanocomplexes were delivered directly to the liver cancer in a rat model by transarterial methods. Anticancer effect was confirmed by magnetic resonance imaging (MRI), and the immune cell population and functional changes were confirmed by flow cytometry (FACS). We first established a rat liver cancer model by implanting McA-RH7777 into rats and confirmed the formation of liver cancer through MRI, pathological examinations, and biochemical tests. Transarterial injection of pPBA-Dox nanocomplexes had a stronger anticancer effect than conventional Dox alone. Higher numbers of CD8+ and CD4+ T cells with activated phenotypes were infiltrated into the tumor microenvironment after transarterial pPBA-Dox treatments than after Dox alone treatment, suggesting the induction of stronger local immune responses by pPBA-Dox than Dox alone. This study provides a theoretical basis for TAC combined with ICIs and insight into novel targeted therapies using nanocomplexes for the treatment of HCC.

The effect of resistance exercise on the immune cell function in humans: A systematic review.

Resistance exercise is beneficial for the immune system, including decreased susceptibility to infections and improved effectiveness of vaccinations. This review aims to provide a systematic analysis of the literature regarding the impact of resistance exercise on immune cells in the blood circulation. The protocol of this review followed the PRISMA guidelines and registered in PROSPERO (ID: CRD42020157834). PubMed and Web-of-Science were systematically searched for relevant articles. Outcomes were divided into two categories: 1) inflammatory gene expression or secretion of inflammation-related cytokines and 2) other aspects such as cell migration, proliferation, apoptosis, phagocytosis, and redox status. Thirty intervention studies were included in this review, of which 11 articles were randomized controlled trials and six non-randomized controlled trials. Although only resistance exercise interventions were included, there was a high heterogeneity regarding specific exercise modalities. The most frequently studied outcome measures were the gene and protein expression levels in peripheral blood mononuclear cells (PBMC). This review reveals that already one acute exercise bout activates the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway in PBMC. Although resistance exercise induces an acute cytosolic oxidative stress response, the antioxidant enzyme expression is improved after resistance training period. Natural killer cell activity increases in older but decreases in younger adults immediately after a resistance exercise bout. Moreover, resistance exercise improves neutrophil phagocytic activity. Finally, effects on lymphocyte proliferation remain unclear. The results of this systematic review demonstrate that resistance exercise has beneficial effects on several aspects of immune cell function both in young and older individuals. Acute changes in immune cell function occur already after a single bout of resistance exercise. However, regular resistance training during several weeks seems necessary to obtain beneficial adaptations that can be related to better immunity and reduced inflammation. The effects documented in this review confirm the beneficial effects of resistance exercise in young as well as older persons on the immune cell function.

Dynamic Immune Function Changes Before and After the First Radioactive Iodine Therapy After Total Resection of Differentiated Thyroid Carcinoma.

The effects of total thyroidectomy or radioactive iodine therapy on immune activation and suppression of the tumor microenvironment remain unknown. We aimed to investigate the effects of these treatments on the immune function in patients with differentiated thyroid carcinoma (DTC). Our cohort included 45 patients with DTC treated with total thyroidectomy and radioactive iodine therapy (RAIT). Immune function tests were performed by flow cytometry at 0, 30, and 90 days post-RAIT. Both the percentage and absolute number of circulating regulatory T cells were significantly lower in the postoperative DTC compared to the healthy controls. Notably, the absolute number of multiple lymphocyte subgroups significantly decreased at 30 days post-RAIT compared to those pre-RAIT. The absolute counts of these lymphocytes were recovered at 90 days post-RAIT, but not at pre-RAIT levels. Additionally, the Th17 cell percentage before RAIT was positively correlated with thyroglobulin (Tg) levels after RAIT. The tumor burden might contribute to increased levels of circulating Tregs. In conclusion, RAIT caused transient radiation damage in patients with DTC and the percentage of Th17 cells before RAIT could be a significant predictor of poor prognosis in patients with DTC.

Immunosafety evaluation in Juvenile Göttingen Minipigs

Although an extrapolation from the clinical experience in adults can often be considered to support the pediatric use for most pharmaceutical compounds, differences in safety profiles between adult and pediatric patients can be observed. The developing immune system may be affected due to exaggerated pharmacological or non-expected effects of a new drug. Toxicology studies in juvenile animals could therefore be required to better evaluate the safety profile of any new pharmaceutical compound targeting the pediatric population. The Göttingen minipig is now considered a useful non-rodent species for non-clinical safety testing of human pharmaceuticals. However, knowledge on the developing immune system in juvenile minipigs is still limited. The objective of the work reported here was to evaluate across-age proportions of main immune cells circulating in blood or residing in lymphoid organs (thymus, spleen, lymph nodes) in Göttingen Minipigs. In parallel, the main immune cell populations from healthy and immunocompromised piglets were compared following treatment with cyclosporin A (CsA) at 10 mg/kg/day for 4 wk until weaning. The study also assessed functionality of immune responses using an in-vivo model after “Keyhole limpet hemocyanin” (KLH) immunization and an ex-vivo lymph proliferation assay after stimulation with Concanavalin A. The results demonstrated variations across age in circulating immune cell populations including CD21+ B-cells, αβ-T- and γδ-T-cells, NK cells, and monocytes. CsA-induced changes in immune functions were only partially recovered by 5 mo after the end of treatment, whereas the immune cell populations affected by the treatment returned to normal levels in animals of the same age. Taken together, the study here shows that in this model, the immune function endpoints were more sensitive than the immunophenotyping endpoints.

Machine Learning Screens Potential Drugs Targeting a Prognostic Gene Signature Associated With Proliferation in Hepatocellular Carcinoma.

Background: This study aimed to screen potential drugs targeting a new prognostic gene signature associated with proliferation in hepatocellular carcinoma (HCC). Methods: CRISPR Library and TCGA datasets were used to explore differentially expressed genes (DEGs) related to the proliferation of HCC cells. Differential gene expression analysis, univariate COX regression analysis, random forest algorithm and multiple combinatorial screening were used to construct a prognostic gene signature. Then the predictive power of the gene signature was validated in the TCGA and ICGC datasets. Furthermore, potential drugs targeting this gene signature were screened. Results: A total of 640 DEGs related to HCC proliferation were identified. Using univariate Cox analysis and random forest algorithm, 10 hub genes were screened. Subsequently, using multiplex combinatorial screening, five hub genes (FARSB, NOP58, CCT4, DHX37 and YARS) were identified. Taking the median risk score as a cutoff value, HCC patients were divided into high- and low-risk groups. Kaplan-Meier analysis performed in the training set showed that the overall survival of the high-risk group was worse than that of the low-risk group (p < 0.001). The ROC curve showed a good predictive efficiency of the risk score (AUC > 0.699). The risk score was related to gene mutation, cancer cell stemness and immune function changes. Prediction of immunotherapy suggetsted the IC50s of immune checkpoint inhibitors including A-443654, ABT-888, AG-014699, ATRA, AUY-922, and AZ-628 in the high-risk group were lower than those in the low-risk group, while the IC50s of AMG-706, A-770041, AICAR, AKT inhibitor VIII, Axitinib, and AZD-0530 in the high-risk group were higher than those in the low-risk group. Drug sensitivity analysis indicated that FARSB was positively correlated with Hydroxyurea, Vorinostat, Nelarabine, and Lomustine, while negatively correlated with JNJ-42756493. DHX37 was positively correlated with Raltitrexed, Cytarabine, Cisplatin, Tiotepa, and Triethylene Melamine. YARS was positively correlated with Axitinib, Fluphenazine and Megestrol acetate. NOP58 was positively correlated with Vorinostat and 6-thioguanine. CCT4 was positively correlated with Nerabine. Conclusion: The five-gene signature associated with proliferation can be used for survival prediction and risk stratification for HCC patients. Potential drugs targeting this gene signature deserve further attention in the treatment of HCC.

The influence of age and sex in sarcoidosis.

This review aims to describe how the clinical manifestations of sarcoidosis may be shaped by the effects of sex hormones and by age dependent changes in immune functions and physiology This review is intended to highlight the need to consider the effects of sex and sex in future studies of sarcoidosis. The clinical manifestations of sarcoidosis differ based on sex and gender There is emerging evidence that female and male hormones and X-linked genes are important determinants of immune responses to environmental antigens, which has important implications for granuloma formation in the context of sarcoidosis Furthermore, sex hormone levels predictably change throughout adolescence and adulthood, and this occurs in parallel with the onset immune senescence and changes in physiology with advanced age. Recent studies indicate that sex and age are important variables shaping the immune response of humans to environmental antigens We posit herein that sex and age are important determinants of sarcoidosis clinical phenotypes Many gaps in our understanding of the roles played by sex and gender in sarcoidosis, and these need to be considered in future studies.

Abstract 6102: Single-cell RNA sequencing of lung adenocarcinoma patients reveals age-dependent tumour microenvironment alterations that facilitate response to immunotherapy

Abstract Background: Immunotherapy has revolutionized the treatment of lung adenocarcinoma (LUAD). While the increase in cancer incidence with age and age-associated changes in immune function have been well characterized, age-related differences in the intratumoral immune populations and how this affects immunotherapy response remain unknown. Methods: We collected clinical data from 3,544 samples treated with immune checkpoint blockade (ICB) at the pan-cancer level. Single-cell RNA sequencing data from 19 LUAD samples were integrated and analyzed. Cell types were identified based on canonical marker gene expression. Results: Aged individuals responded more efficiently to ICB treatment at the pan-cancer level. By focusing on LUAD, we showed that patients aged above 46 responded better and had longer progression-free survival than younger counterparts. Further, we presented a 58,031-cell catalog of age-associated single-cell transcriptomes from LUAD patients. Our analysis revealed the accumulation of immune response and chronic inflammation with increasing age. Aged tissues (&amp;gt; 60 years) displayed dominant exhausted natural killer (NK)/T cells and T regulatory cells and were enriched with two subtypes of tumour-associated macrophages marked by SPP1 and CX3CR1 expression, respectively. Furthermore, the ageing dynamics of stroma cells created a pro-tumor microenvironment reflected by increased angiogenesis and collagen formation. Conclusions: These results revealed an increasing immune-suppressive tumour microenvironment with age, supporting an observation of improved ICB response in older LUAD patients. Our extensive single-cell analysis enhances the understanding of age-dependent molecular and cellular dynamics in LUAD. Our results highlight the importance of considering age as a factor for ICB response and provide a rich resource for developing therapeutic targets in LUAD-microenvironment interactions. Citation Format: Xiaoqiang Zhu, Hai Wang, Xuefeng Pan, Jie Hong, Jason Wing Hon Wong, Haoyan Chen, Youwei Zhang. Single-cell RNA sequencing of lung adenocarcinoma patients reveals age-dependent tumour microenvironment alterations that facilitate response to immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6102.

Abstract 2537: Beta-glucan reprograms immunomodulatory metabolism in human macrophage and ex vivo in lung cancer tissues

Abstract We combined multiplex Stable Isotope-Resolved Metabolomics (mSIRM) with Reverse Phase Protein Array (RPPA) to map the time course changes of IM metabolic network and key protein regulators in four human donors’ MΦ’s in response to differential polarization and whole glucan particulates (WGP) treatments. We found consistent and plastic network responses to polarization durations and WGP treatments compared to those of the mouse counterparts. Consistent responses included enhanced 15N2-tryptophan catabolism to quinolinate, 2H2-glucose oxidation to ribose/ribulose-5-phosphate, and conversion of 13C5-glutamine to itaconate in response to pro-inflammatory (M1) versus anti-inflammatory (M2) stimuli. WGP robustly induced the buildup of 2H2-glucose-derived 6-phosphogluconate, -lactate, and -IMP, 13C5-glutamine-derived fructose-1,6-bisphosphate, and increased enrichment of 2H-labeled UDP-N-acetylglucosamine in M2-MΦ’s. However, the Krebs cycle activity was variably enhanced by M2 stimuli or WGP treatment. The consistent effects were related to increased release of proinflammatory IM effectors IL-1β/CXCL10/IFNγ/TNFα by M1-MΦ’s and enhanced release of IL-1β/TNFα to above M1-MΦ’s levels in WGP-treated M2-MΦ’s while boosting the latter in anti-inflammatory IL-10 release and maintenance of NAD+ synthesis. They were also related to lower phagocytosis in M1-MΦ’s and WGP-treated M2-MΦ’s versus M2-MΦ’s. Together with the expression changes of key protein regulators, we suggest enhanced tryptophan catabolism with blocked NAD+ and UTP synthesis to be key to the consistent changes in immune functions in response to M1 stimuli. Likewise, increased glucose utilization via glycolysis and the oxidative branch of the pentose phosphate pathway, and blockade of glutamine-fueled N-linked glycosylation could be linked to reversion of M2 to M1-type immune functions. Reprogrammed Krebs cycle and glutamine conversion to UTP occurred variably in WGP-treated ex vivo organotypic tissue cultures (OTCs) of human non-small cell lung cancer (NSCLC), which could reflect variable M1 repolarization of tumor associated MΦ’s. This in turn correlated with IL-1β/TNFα releases and compromised tumor status, making patient-derived OTCs a unique model for studying variable immunotherapeutic efficacy in cancer patients. In conclusion, consistent and variable IM metabolic responses were evident in four human donors’ MΦ’s. WGP repolarized some M2 to M1-type responses while boosting other M2-type responses. NSCLC OTCs from six patients showed variable M1 repolarization in response to WGP. Citation Format: Teresa W. M. Fan, Saeed Daneshmandi, Teresa A. Cassel, Mohammad B. Uddin, James Sledziona, Patrick T. Thompson, Penghui Lin, Richard M. Higashi, Andrew N. Lane. Beta-glucan reprograms immunomodulatory metabolism in human macrophage and ex vivo in lung cancer tissues [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2537.

Abstract CT115: Safety and efficacy of Anti-PD-L1 antibody MEDI4736 (durvalumab) in combination with capecitabine in patients with advanced solid tumors (DURVA+)

Abstract Background: It is hypothesized that chemotherapy alters the tumor microenvironment (TME) by increasing T cell infiltration and enhancing antigen presentation to T cells, and thereby may enhance immune-mediated cytotoxicity and tumor response to immunotherapy. The goal of this trial (DURVA+) is to evaluate and compare the immune-modulating effects of six different chemotherapy agents with Durvalumab (D), a PD-L1 checkpoint inhibitor. We report here the first analysis of one of the arms of DURVA+: D plus capecitabine (C). Methods: This is a non-randomized phase 2 trial for adult patients (pts) with refractory advanced solid tumors. Pts received C at a dose of 1250 mg/m2 bid on days 1-14 of 21-day cycle. For the first 3 cycles, D is administered every 2 weeks at a dose of 750 mg IV starting on C1D8. From cycle 4 onwards, D was given at a dose of 1125 mg IV every 3 week (day 1 of each 21-day cycle). Tumor biopsies were required at baseline, C1D8 (prior to durvalumab), C3D8, and optional at progression. Radiologic tumor assessment occurs every 4 cycles by RECIST. iRECIST is assessed as a secondary endpoint. Baseline and at least one on-treatment biopsy were obtained for 11 patients, at least 8 of which are evaluable for pharmacodynamic (PD) response (treatment-induced changes in immune cell tumor content and function). Results: As of Nov 10, 2021, 14 pts were enrolled to this arm. Median age 57 (range 31-73 years), 7 males and 7 females, 4 black, 8 white, 2 unknown race. Majority of pts had colorectal carcinoma (ca) (n=8), other include (n=1 each): appendiceal, anal squamous cell ca, leiomyosarcoma, pancreatic ca, breast ca, and nasopharyngeal squamous cell ca. Median lines of prior treatment was 3.5 (1-13), 4 pts previously received ICI. Median number of days on study treatment were 83 days. One pt experience partial response (breast) and five pts experienced SD (3 colon, 1 pancreatic, 1 non-uterine leiomyosarcoma) for a median duration of 136 days as of Jan 3, 2022 (data cutoff); one pt with pancreatic ca experienced a 27% reduction in sum of target lesions. Treatment related adverse events (TrAE) of all grades occurred in 13 pts, serious TrAEs (more than grade 2) occurred in 8 pts and included anemia, pulmonary embolism, hyperbilirubinemia, palmar plantar syndrome, neutropenia, lymphopenia and diarrhea. Immune-mediated AEs included transaminitis/hepatitis (n=4, all grade 1/2), hypothyroidism (n=2, grade 1/2), rash (n=1, grade 1) and colitis (grade 3). PD studies are still ongoing. Conclusion: The combination of durvalumab with capecitabine is feasible with no unexpected adverse events. PD assessment is ongoing to examine the hypothesis that chemotherapy alters tumor micronenvironment (TME) and T cell activation. The study was sponsored by NCI, and was funded in part by NCI Contract HHSN261200800001E. DURVA+ is registered with ClinicalTrials.gov, NCT03907475. Citation Format: Mohamad Adham Salkeni, Geraldine O’Sullivan Coyne, Naoko Takebe, Elad Sharon, Howard Streicher, Jessica Mukherjee, Ashley Bruns, Arjun Mitra, Abdul R. Naqash, Larry Rubinstein, Kristin K. Fino, King L. Fung, Katherine Ferry-Galow, Ralph Parchment, Helen Chen, James Doroshow, Alice Chen. Safety and efficacy of Anti-PD-L1 antibody MEDI4736 (durvalumab) in combination with capecitabine in patients with advanced solid tumors (DURVA+) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT115.

Sex-specific effects of daily tadalafil on diabetic heart kinetics in RECOGITO, a randomized, double-blind, placebo-controlled trial.

Cyclic GMP-phosphodiesterase type 5 (PDE5) inhibition has been shown to counteract maladaptive cardiac changes triggered by diabetes in some but not all studies. We performed a single-center, 20-week, double-blind, randomized, placebo-controlled trial (NCT01803828) to assess sex differences in cardiac remodeling after PDE5 inhibition in patients with diabetic cardiomyopathy. A total of 122 men and women (45 to 80 years) with long-duration (>3 years) and well-controlled type 2 diabetes mellitus (T2DM; HbA1c < 86 mmol/mol) were selected according to echocardiographic signs of cardiac remodeling. Patients were randomly assigned (1:1) to placebo or oral tadalafil (20 mg, once daily). The primary outcome was to evaluate sex differences in cardiac torsion change. Secondary outcomes were changes in cardiovascular, metabolic, immune, and renal function. At 20 weeks, the treatment-by-sex interaction documented an improvement in cardiac torsion (-3.40°, -5.96; -0.84, P = 0.011) and fiber shortening (-1.19%, -2.24; -0.14, P = 0.027) in men but not women. The primary outcome could not be explained by differences in cGMP concentrations or tadalafil pharmacodynamics. In both sexes, tadalafil improved hsa-miR-199-5p expression, biomarkers of cardiovascular remodeling, albuminuria, renal artery resistive index, and circulating Klotho concentrations. Immune cell profiling revealed an improvement in low-grade chronic inflammation: Classic CD14++CD16- monocytes reduced, and Tie2+ monocytes increased. Nine patients (14.5%) had minor adverse reactions after tadalafil administration. Continuous PDE5 inhibition could offer a strategy to target cardiorenal complications of T2DM, with sex- and tissue-specific responses. Further studies are needed to confirm Klotho and hsa-miR-199-5p as markers for T2DM complications.

Short-term adaptability to non-hyperthermal stress: Antioxidant, immune and gut microbial responses in the red swamp crayfish, Procambarus clarkii

Thermal stress is a common experience suffered in rice-crayfish integrated systems (RCIS) by the red swamp crayfish, Procambarus clarkii. Based on our previous studies, the adaptability of this crayfish to non-hyperthermal stress over a specified period was explored within a short term by investigating the antioxidant capacity, immune function, HSP70 expression and intestinal microbial changes. Under the non-hyperthermal stress temperature (30 °C) determined by cumulative survivals, the malondialdehyde (MDA) content, total antioxidant capacity (T-AOC), total super oxide dismutase (T-SOD) and catalase (CAT) activities in hepatopancreas significantly increased to the peak values during at 12 h or 24 h (P < 0.05), respectively, and then gradually recovered to the levels of the controls (24 °C) (P > 0.05); the immune-related enzymatic activities (alkaline phosphatase, AKP; acid phosphatase, ACP; lysozyme, LZM) and gene expressions (TNFAIP, TNF and Toll) rapidly increased and reach the peaks at 12 h and 48 h, respectively, and maintained subsequently the higher levels compared with the controls (P < 0.05) except for TNFAIP. The relative expressions of HSP70 in the hepatopancreas, intestine and gill all increased sharply and reached the peak at 6 h (P < 0.05), then decreased fast and roughly returned to the levels of the controls at 96 h (P > 0.05), while, there was still a relatively high expression during the later stages in the gill. High-throughput sequencing of 16S ribosomal DNA revealed the thermal stress altered the diversity and composition of intestinal microbiota. The increased abundance of Proteobacteria and decreased abundance of Bacteroidetes (mainly Bacteroides) and Firmicutes might be indicative of the elevated microbial instability and the unnecessity for high level of nutrient metabolism in the host. The abundance increase of relevant metabolic pathways (e.g., the biosynthesis and degradation of amino acids and lipids) predicted by MetaCyc might be caused by the abundance increase of Proteobacteria. The decreased abundance of two phyla (Actinobacteria and Fusobacteria) and genus Anaerorhabdus could adversely affect the crayfish's intestinal health status. While, the obvious abundance decrease of putatively opportunistic genera (Shewanella and Acinetobacter), combined with the abundance increase of beneficial phylum Tenericutes and genus Rhodobacter, suggested that there concurrently existed some adaptability to the non-thermal stress, which did not resulted in apparent risks of intestinal disorder. This study provides insights into the underling adaptive mechanisms associated with the anti-oxidation, immunity, intestinal microflora and nutrient metabolism of P. clarkii in response to the short-term non-hyperthermal stress.