Severe Body Weight Loss Research Articles

Sight of Aged Microplastics Adsorbing Heavy Metal Exacerbated Intestinal Injury: A Mechanistic Study of Autophagy-Mediated Toxicity Response.

Contaminant-bearing polystyrene microplastics (PSMPs) may exert significantly different toxicity profiles from their contaminant-free counterparts, with the role of PSMPs in promoting contaminant uptake being recognized. However, studies investigating the environmentally relevant exposure and toxic mechanisms of aged PSMPs binding to Cr are limited. Here, we show that loading of chromium (Cr) markedly alters the physicochemical properties and toxicological profiles of aged PSMPs. Specifically, Cr-bearing aged PSMPs induced severe body weight loss, oxidative stress (OS), autophagy, intestinal barrier injury, inflammation-pyroptosis response, and enteropathogen invasion in mice. Mechanistic investigations revealed that PSMPs@Cr exacerbated the OS, resulting in intestinal barrier damage and inflammation-pyroptosis response via overactivated Notch signaling and autophagy/cathepsin B/IL-1β pathway, respectively, which ultimately elevated mortality related to bacterial pathogen infection. In vitro experiments confirmed that autophagy-mediated reactive oxygen species (ROS) overproduction resulted in severe pyroptosis and impaired intestinal stem cells differentiation alongside the overactivation of Notch signaling in PSMPs@Cr-exposed organoids. Overall, our findings provide an insight into autophagy-modulated ROS overproduction within the acidic environment of autophagosomes, accelerating the release of free Cr from PSMPs@Cr and inducing secondary OS, revealing that PSMPs@Cr is a stable hazard material that induces intestinal injury. These findings provided a potential therapeutic target for environmental MPs pollution caused intestinal disease in patients.

Gut microbiota and their relationship with circulating adipokines in an acute hepatic encephalopathy mouse model induced by surgical bile duct ligation

Background and aimsVarious studies have shown the importance of the gut microbiota in human health. However, little is known about gut microbiome patterns and their effect on circulating adipo-myokine levels in hepatic encephalopathy (HE). We investigated the relationship between the gut microbiota and adipo-myokine levels using a mouse model of HE induced by surgical bile duct ligation (BDL). Methods and resultsWild-type C57BL/6J mice were subjected to sham surgery or BDL. Severe body weight loss, suppressed feed intake, and liver failure were observed in BDL mice compared with sham control mice. Additionally, changes in gut microbial communities and serum adipo-myokine levels were noted in BDL mice. In the BDL mouse gut, we identified 15 differentially abundant taxa including the phylum Verrucomicrobiota, the classes Actinomycetes and Verrucomicrobiae, the order Verrucomicrobiales, the families Akkermansiaceae, Bacteroidaceae, Rikenellaceae, and Oscillospiraceae, the genera Alistipes, Akkermansia, Muribaculum, and Phocaeicola, and the species Akkermansia muciniphila, Alistipes okayasuensis, and Muribaculum gordoncarteri by LEfSe analysis (LDA score≥4.0). Higher levels of certain adipo-myokines such as BDNF were detected in the serum of BDL mice. Spearman correlation analysis revealed that certain adipo-myokines (e.g., FSTL1) were positively correlated with the class Actinomycetes, the family Rikenellaceae, the genus Alistipes, and the species Alistipes okayasuensis. Interestingly, A. okayasuensis and M. gordoncarteri, recently isolated microbes, showed richness in the gut of BDL mice and demonstrated positive correlations with adipo-myokines such as FGF21. ConclusionsOverall, our results suggest that alteration of the gut microbiota in patients with HE may be closely correlated to the levels of adipo-myokines in the blood.

Clinical Significance of Body Weight Loss During Chemotherapy for Advanced Gastric Cancer Undergoing Conversion Surgery.

The purpose of the present study was to assess the clinical impact of body weight loss (BWL) during chemotherapy in patients with initially unresectable advanced gastric cancer who underwent conversion surgery. This retrospective study included 61 patients with stage IV gastric cancer who underwent conversion surgery after chemotherapy, and body weight changes during chemotherapy were examined. Based on receiver operating characteristic (ROC) curve analysis of body weight change for disease recurrence, the cutoff value of BWL was determined. Based on the BWL cutoff value, patients were classified into two groups. Body weight change ranged from 28.2% to -21.8%. The cut-off value of BWL was set at 6% based on the ROC analysis. Of the 61 patients, 45 (74%) and 16 (26%) had <6% and ≥6% BWL, respectively. Patients with ≥6% BWL had peritoneal dissemination, pathological lymph node metastasis, residual tumor status of R1-2, and disease recurrence compared with those with <6% BWL (all p<0.05). The median survival times after conversion surgery were 21 and 63 months in the ≥6% and <6% BWL groups, respectively (p<0.01). Univariate analysis identified BWL as an independent prognostic factor (p=0.01), although histological response alone was significantly associated with survival in the multivariate analysis (p=0.02). Patients with severe BWL during chemotherapy may be excluded from the indication of conversion surgery.

Molecular Iodine Improves the Efficacy and Reduces the Side Effects of Metronomic Cyclophosphamide Treatment against Mammary Cancer Progression.

Metronomic chemotherapy with cyclophosphamide (Cpp) has shown promising results in cancer protocols. These lower and prolonged doses have antiangiogenic, pro-cytotoxic, and moderate secondary effects. Molecular iodine (I2) reduces the viability of cancer cells and, with chemotherapeutic agents, activates the antitumoral immune response and diminishes side effects. The present work evaluates the adjuvant of oral I2 with Cpp using a murine model of mammary cancer. Female Sprague Dawley rats with 7,12-dimethylbenzantracene-induced tumors received Cpp intraperitoneal (50 and 70 mg/kg two times/week, iCpp50 and iCpp70) and oral (0.03%; 50 mg/Kg; oCpp50) doses. I2 (0.05%, 50 mg/100 mL) and oCpp50 were offered in drinking water for three weeks. iCpp70 was the most efficient antitumoral dose but generated severe body weight loss and hemorrhagic cystitis (HC). I2 prevented body weight loss, exhibited adjuvant actions with Cpp, decreasing tumor growth, and canceled HC mechanisms, including decreases in vascular endothelial growth factor (VEGF) and Survivin expression. oCpp50 + I2 diminished angiogenic signals (CD34, vessel-length, and VEGF content) and proinflammatory cytokines (interleukin-10 and tumor necrosis factor-alpha) and increased cytotoxic (lymphocytic infiltration, CD8+ cells, Tbet, and interferon-gamma) and antioxidant markers (nuclear erythroid factor-2 and glutathione peroxidase). I2 enhances the effectiveness of oCpp, making it a compelling candidate for a clinical protocol.

Seasonal and fasting induced changes in iron metabolism in Djungarian hamsters

Djungarian hamsters are small rodents that show pronounced physiological acclimations in response to changes in photoperiod, and unfavorable environmental conditions such as reduced food availability and low external temperature. These include substantial adjustments, such as severe body weight loss and the use of daily torpor. Torpor is a state of decreased physiological activity in eutherms, usually marked by low metabolic rate and a reduced body temperature. In this study, we investigated the effects of photoperiodic acclimation and food deprivation on systemic iron metabolism in Djungarian hamsters. Our study illustrates the association between liver iron levels and the incidence of torpor expression during the course of the experiment. Moreover, we show that both, acclimation to short photoperiods and long-term food restriction, associated with iron sequestration in the liver. This effect was accompanied with hypoferremia and mild reduction in the expression of principal iron-hormone, hepcidin. In addition to iron, the levels of manganese, selenium, and zinc were increased in the liver of hamsters under food restriction. These findings may be important factors for regulating physiological processes in hamsters, since iron and other trace elements are essential for many metabolic and physiological processes.

Abstract P165: Hypertension Increases Susceptibility To Experimental Malaria In Mice

Despite available treatments, global prevalence of hypertension is on the rise and burdening healthcare, especially in under-developed countries where infectious diseases are also rampant. Previously we reported that hypertension is associated with abnormal red blood cell (RBC) physiology and anemia. Since RBCs are target host cells for malarial pathogen, Plasmodium , we hypothesized that hypertensive patients with abnormal RBC physiology are at an increased risk of susceptibility to Plasmodial infection. Methods: To test this hypothesis, eight weeks old hypertensive BPH/2J and normotensive BPN/3J female mice (n=6) were first characterized for their RBC physiology and subsequently infected with Plasmodium yoelii (P. yoelii) , a murine-specific non-lethal strain. Results: Compared to BPN mice, BPH mice displayed comparable RBC count, but lower hemoglobin (BPH 16.3±0.8, vs BPN 16.5±1.0 mg/dL; p&lt;0.01); lower % hematocrit (BPH 59.6 ±2.4 vs BPN 61.0 ± 2.7; p&lt;0.01), and lower mean corpuscular volume (BPH 52.6±3.13 vs BPN 54.2±1.6 fl; p&lt;0.005), indicating microcystic anemia. Further, RBCs from BPH mice were resistant to osmotic hemolysis in hypotonic (0.42%) NaCl solution (% hemolysis BPH 43.2±1.6% vs 97.3±1.8% BPN; p&lt;0.001). Lower membrane fluidity was observed in the RBCs of BPH mice as noted through higher levels of the saturated fatty acid, stearate (BPH 16.2±0.1% vs BPN 13.9±0.16%; p&lt;0.001), phospholipids (BPH 554±18.3 vs BPN 496±6.2 μg/mg protein p=0.02) and lower polyunsaturated fatty acids like arachidonic acid (BPH 13.7±0.1% vs BPN 15.1±0.5%, BPN; p=0.03). Further, upon infection for 7 days with P. yoelii , BPH mice experienced severe body weight loss compared to minimal change in body weight noted in the BPN mice (BPH Day 0 18.53±0.32 g vs Day 7 16.43±0.71 g vs BPN Day 0 22.5±1.28 g vs Day 7 22.43±1.30 g; p&lt;0.01). This was accompanied by a significantly decreased number of RBC compared to BPN mice (BPH 3.4±1.3х10 12 vs BPN 5.5±1.2х10 12 cells/L; p=0.03), indicating that BPH mice were more susceptible to malaria infection. Conclusion: Collectively, these data demonstrate that aberrant RBC physiology observed in hypertensive mice contributes to an increased susceptibility to experimental malaria.

Skin Wound following Irradiation Aggravates Radiation-Induced Brain Injury in a Mouse Model.

Radiation injury- and radiation combined with skin injury-induced inflammatory responses in the mouse brain were evaluated in this study. Female B6D2F1/J mice were subjected to a sham, a skin wound (SW), 9.5 Gy 60Co total-body gamma irradiation (RI), or 9.5 Gy RI combined with a skin puncture wound (RCI). Survival, body weight, and wound healing were tracked for 30 days, and mouse brain samples were collected on day 30 after SW, RI, RCI, and the sham control. Our results showed that RCI caused more severe animal death and body weight loss compared with RI, and skin wound healing was significantly delayed by RCI compared to SW. RCI and RI increased the chemokines Eotaxin, IP-10, MIG, 6Ckine/Exodus2, MCP-5, and TIMP-1 in the brain compared to SW and the sham control mice, and the Western blot results showed that IP-10 and p21 were significantly upregulated in brain cells post-RI or -RCI. RI and RCI activated both astrocytes and endothelial cells in the mouse brain, subsequently inducing blood-brain barrier (BBB) leakage, as shown by the increased ICAM1 and GFAP proteins in the brain and GFAP in the serum. The Doublecortin (DCX) protein, the "gold standard" for measuring neurogenesis, was significantly downregulated by RI and RCI compared with the sham group. Furthermore, RI and RCI decreased the expression of the neural stem cell marker E-cadherin, the intermediate progenitor marker MASH1, the immature neuron cell marker NeuroD1, and the mature neuron cell marker NeuN, indicating neural cell damage in all development stages after RI and RCI. Immunohistochemistry (IHC) staining further confirmed the significant loss of neural cells in RCI. Our data demonstrated that RI and RCI induced brain injury through inflammatory pathways, and RCI exacerbated neural cell damage more than RI.

Abstract 2006: A novel macromolecular platinum drug with potent efficacy to overcome cross-resistance in cancer therapy

Abstract Background: Small molecular platinum (Pt)-based drugs such as cisplatin, carboplatin, and oxaliplatin have achieved great success in clinic over 40 years in cancer therapy. However, lack of tumor specificity leads to severe side effects. In addition, cancer cells frequently develop resistance to treatment through overexpressing cell surface multi-drug resistant proteins (MDRs). Therefore, it is desirable to develop novel Pt-based drugs with excellent anti-cancer efficacy and a minimal toxicity profile, as well as the capability to overcome drug resistance. We have previously demonstrated that poly(amino acid)-conjugated drugs can reduce toxicity and improve pharmacokinetics of chemotherapeutics and also overcome drug resistance by bypassing MDRs. Here we would like to apply this strategy on Pt drug development. Method: To address this question, we synthesized a L-aspartic acid-chelated Pt moieties (Asp-Pt), and then conjugated it to poly(L-glutamic acid/L-aspartic acid) to generate a polymeric-Pt macromolecule, namely carrier-platin. We next applied this drug on cancer cell lines including the therapeutic-resistant cancer cells in vitro, and tumor bearing murine models in vivo to evaluate its efficacy and toxicity. Result: We first tested the cytotoxicity of carrier-platin and compared with a current first-line chemotherapeutic oxaliplatin on colorectal cancer cell CT26 in vitro. Surprisingly, we found that carrier-platin effectively killed the CT26 cancer cells with hours while oxaliplatin slowly induced cell death after days. Importantly, carrier-platin was well-tolerated by non-cancerous cells. Furthermore, carrier-platin killed cancer cells that resistant to current chemotherapeutics in clinic, including cisplatin, oxaliplatin, doxorubicin and docetaxel, as effective as their parental cells. Next, we found that carrier-platin showed prolonged circulation time and high tumor accumulation compared with Pt-equivalent dosage of oxaliplatin. Treatment with 8 mg Pt/kg oxaliplatin caused severe body weight loss and intolerable hematotoxicity in mice, but even higher dose of carrier-platin (12 mg Pt/kg) can be tolerated by mice with stable body weight and did not induce any clinical symptoms. Finally, we revealed that carrier-platin potently inhibited mouse colorectal (CT26) and human ovarian (A2780) tumor growth over Pt-equivalent dosage of oxaliplatin. More importantly, carrier-platin was almost as effective on the cisplatin-resistant A2780 tumors (A2780-Cis) as on the parental A2780 tumors in vivo. Conclusion: We developed a novel macromolecular drug carrier-platin that overcome shortages of current small molecular Pt drugs. It displayed a satisfactory toxicity profile accompanied with excellent anti-tumor efficacy in vivo. Moreover, carrier-platin overcomes the cross-resistance problem of cancer cells to current chemotherapeutic drugs. Citation Format: Yongbin Liu, Dongfang Yu, Junhua Mai, Ping-Ying Pan, Shu-Hsia Chen, Haifa Shen. A novel macromolecular platinum drug with potent efficacy to overcome cross-resistance in cancer 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 2006.

Construction of Activity-based Anorexia Mouse Models.

Anorexia nervosa (AN) is a psychiatric disorder mainly characterized by extreme hypophagia, severe body weight loss, hyperactivity, and hypothermia. Currently, AN has the highest mortality rate among psychiatric illnesses. Despite decades of research, there is no effective cure for AN nor is there a clear understanding of its etiology. Since a complex interaction between genetic, environmental, social, and cultural factors underlines this disorder, the development of a suitable animal model has been difficult so far. Here, we present our protocol that couples a loss-of-function mouse model to the activity-based anorexia model (ABA), which involves self-imposed starvation in response to exposure to food restriction and exercise. We provide insights into a neural circuit that drives survival in AN and, in contrast to previous protocols, propose a model that mimics the conditions that mainly promote AN in humans, such as increased incidence during adolescence, onset preceded by negative energy balance, and increased compulsive exercise. This protocol will be useful for future studies that aim to identify neuronal populations or brain circuits that promote the onset or long-term maintenance of this devastating eating disorder.

Increased sensitivity to chemically induced colitis in mice harboring a DNA-binding deficient aryl hydrocarbon receptor.

The aryl hydrocarbon receptor (AHR), a transcription factor best known for mediating toxic responses of environmental pollutants, also integrates metabolic signals to promote anti-inflammatory responses, intestinal homeostasis, and maintain barrier integrity. AHR regulates its target genes through direct DNA-binding to aryl hydrocarbon response elements (AHREs) but also through tethering to other transcription factors in a DNA-binding independent manner. However, it is not known if AHR's anti-inflammatory role in the gut requires its ability to bind to AHREs. To test this, we determined the sensitivity of Ahrdbd/dbd mice, a genetically modified mouse line that express an AHR protein incapable of binding to AHREs, to dextran sulfate sodium (DSS)-induced colitis. Ahrdbd/dbd mice exhibited more severe symptoms of intestinal inflammation than Ahr+/+ mice. None of the Ahrdbd/dbd mice survived after the 5-day DSS followed by 7-day washout period. By day 6, the Ahrdbd/dbd mice had severe body weight loss, shortening of the colon, higher disease index scores, enlarged spleens, and increased expression of several inflammation genes, including interleukin 1b (Il-1b), Il-6, Il-17, C-x-c motif chemokine ligand 1 (Cxcl1), Cxcl2, Prostaglandin-endoperoxide synthase (Ptgs2), and lipocalin-2. Our findings show that AHR's DNA-binding domain and ability to bind to AHREs are required to reduce inflammation, maintain a healthy intestinal environment, and protect against DSS-induced colitis.

Microplastics perturb colonic epithelial homeostasis associated with intestinal overproliferation, exacerbating the severity of colitis

A great amount of the population died due to living or working in an unhealthy environment, highlighting the critical role of environmental pollutants in inducing diseases. Microplastics are widespread environmental pollutants and have been found in various tissues of human beings, yet the risk of microplastics in the occurrence of disease, especially environmentally-related colitis, is unclear. This study focused on the effects of microplastics exposure on intestinal homeostasis and the initiation of colitis. We noticed that microplastics exposure had a limited impact on mice, as verified by no difference observed in bodyweight change, IL-1β and IL-6 levels in jejunum and liver. Nevertheless, in the colon, the IL-1β and IL-6 levels were slightly increased and the goblet cell number was decreased. Interestingly, we observed that crypt number and depth, the levels of intestinal stem cell markers, combined with the expression of proliferating cell nuclear antigen and proto-oncogene c-Myc were all significantly increased with microplastics treatment, indicating the overproliferation of colonic mucosa. The effect of microplastics on proliferation and differentiation of crypt was further demonstrated to be regulated by the overactivation of the Notch signaling pathway in intestinal organoids. Furthermore, microplastics exposure accelerated the development of colitis with severe bodyweight loss, diarrhea and bloody stools, macroscopic and pathological damage, and inflammation levels. Worsened liver pathological damage and inflammation in mice with colitis under microplastics exposure also were found. These results suggested that microplastics disrupted the balance between colonic epithelium self-renewal and differentiation, exacerbating the colitis, and might be an environmental-related disease risk factor.

Update on the Etiology, Assessment, and Management of COPD Cachexia: Considerations for the Clinician.

Cachexia is a commonly observed but frequently neglected extra-pulmonary manifestation in patients with chronic obstructive pulmonary disease (COPD). Cachexia is a multifactorial syndrome characterized by severe loss of body weight, muscle, and fat, as well as increased protein catabolism. COPD cachexia places a high burden on patients (eg, increased mortality risk and disease burden, reduced exercise capacity and quality of life) and the healthcare system (eg, increased number, length, and cost of hospitalizations). The etiology of COPD cachexia involves a complex interplay of non-modifiable and modifiable factors (eg, smoking, hypoxemia, hypercapnia, physical inactivity, energy imbalance, and exacerbations). Addressing these modifiable factors is needed to prevent and treat COPD cachexia. Oral nutritional supplementation combined with exercise training should be the primary multimodal treatment approach. Adding a pharmacological agent might be considered in some, but not all, patients with COPD cachexia. Clinicians and researchers should use longitudinal measures (eg, weight loss, muscle mass loss) instead of cross-sectional measures (eg, low body mass index or fat-free mass index) where possible to evaluate patients with COPD cachexia. Lastly, in future research, more detailed phenotyping of cachectic patients to enable a better comparison of included patients between studies, prospective longitudinal studies, and more focus on the impact of exacerbations and the role of biomarkers in COPD cachexia, are highly recommended.

Effect of Patient-Participation Continuous Nutritional Counseling in Gastric Cancer Patients who Underwent Gastrectomy.

Body weight loss (BWL) and skeletal muscle loss (SML) are inevitable after gastrectomy for gastric cancer (GC) and can decrease patients' quality of life (QOL) and survival. The aim of this retrospective study was to evaluate the effect of perioperative and post-discharge patient participation in continuous nutritional counseling (CNC) on post-gastrectomy BWL and SML. Ninety-three patients with GC who underwent curative gastrectomy between March 2018 and July 2019 were analyzed. Patients received either pre-discharge nutritional counseling alone (control group, n = 49) or patient-participation CNC (CNC group, n = 44) after gastrectomy. Differences between percentage BWL (%BWL), percentage SML (%SML), and nutrition-related blood parameters between the preoperative values and those at 12 months after surgery were compared between the groups. Compared with the control group, %BWL was significantly lower in the CNC group at 1 month (-6.2 ± 2.5% vs. -7.9 ± 3.3%, p = 0.005), 6 months (-7.8 ± 6.6% vs. -12.3 ± 6.4%, p = 0.001) and 12 months (-7.9 ± 7.6% vs. -13.2 ± 8.2%, p = 0.002), and %SML was significantly lower in the CNC group at 12 months (-5.3 ± 10.3% vs. -12.8 ± 12%, p = 0.002). Regarding nutrition-related blood parameters, change in total cholesterol was significantly lower in the CNC group than the control group at 12 months after surgery (p = 0.02). Multivariate analysis identified no CNC as an independent risk factor for severe BWL (p = 0.001) and SML (p = 0.006) at 12 months after surgery. Following gastrectomy, patient-participation CNC prevented postoperative BWL and SML after surgery. These results support the induction of such a CNC program in these patients.

Abstract 1865: Tumor microenvironment ameliorating effect of E7130 sensitized ER+ breast cancer against fulvestrant

Abstract Introduction and objectives: E7130 is a novel anti-cancer agent created from a total synthetic study of the natural compound norhalichondrin B, and is evaluated in a clinical study currently (NCT03444701). Previous pre-clinical study reported that E7130 had both cancer cell proliferation inhibitory activity and tumor microenvironment ameliorating activities featuring vascular remodeling effect and anti-CAF (cancer-associated fibroblasts) effect. Intratumoral hypoxia induces tumor malignancy and causes resistance against anti-cancer drugs including anti-estrogen agents (eg fulvestrant). It is hypothesized that vascular remodeling effect of E7130 mitigates hypoxia-induced resistant mechanism of ER (estrogen receptor) + breast cancer against anti-estrogen agents, resulting in improving anti-tumor activity of fulvestrant in combination with E7130. Experimental procedure: ERα expression levels in human ER+ breast cancer cell lines under hypoxic or normoxic culture condition were examined by FCM (flow cytometry) analysis. The histological changes after treatment with E7130 were examined by immunohistochemical analyses in the MCF-7 subcutaneous xenograft model. Anti-tumor activity of E7130 in combination with fulvestrant were examined in concurrent and in sequential treatment in the MCF-7 model. Summary: In vitro hypoxic culture condition down-regulated ERα expression in MCF-7 and T-47D compared with normoxic condition. E7130 had no effect on the ERα expression level in the cancer cells in vitro. On the contrary, E7130 increased ERα expression level in the cancer cells concomitantly with an increase of CD31-positive endothelial cell number (ie vascular remodeling effect) in MCF-7 xenograft tumor. These results suggested that E7130 up-regulated ERα expression in cancer cells not by cell-autonomous manner but by mitigating intratumoral hypoxia via its vascular remodeling effect. Concurrent combination therapy of E7130 at 90 μg/kg with fulvestrant at 250 mg/kg significantly inhibited the tumor growth compared with each monotherapy. In sequential combination therapy, pretreatment of E7130 followed by fulvestrant showed superior anti-tumor activity to posttreatment of E7130 following fulvestrant. Severe body weight loss was not observed in any treatment groups. Conclusions: The vascular remodeling effect of E7130 increased ERα expression in cancer cells in vivo, and sensitized ER+ breast cancer against fulvestrant to achieve promising combinational anti-tumor activity. Our data provides compelling evidence that E7130 attenuates tumor malignancy by its tumor microenvironment ameliorating effects and improves current anti-cancer therapy in combination with other drugs. Citation Format: Takanori Abe, Satoshi Kawano, Ken Ito, Taro Semba, Kimiyo Tabata, Hiroki Jozawa, Osamu Asano, Yoshito Kishi. Tumor microenvironment ameliorating effect of E7130 sensitized ER+ breast cancer against fulvestrant [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 1865.

Abstract 3505: CLN-617 is an IL-2/IL-12 fusion protein with a collagen-anchoring domain that induces potent systemic anti-tumor immunity upon intra-tumoral administration

Abstract Despite strong anti-tumor immune responses in the clinic, cytokine therapy for cancer treatment, has remained limited in use due to its narrow therapeutic window. Moreover, anti-tumor immunity is typically driven through multiple cytokines acting in concert in a local tumor microenvironment (TME) via autocrine or paracrine signaling. Yet most cytokine therapies focus on systemic delivery of individual cytokines causing adverse events without achieving therapeutic doses at the tumor site. Recently, intratumoral (IT) delivery of cytokines such as IL-2 and IL-12 anchored to collagen in the TME has resulted in promising anti-tumor efficacy with minimal toxicity in multiple murine tumor models, and remarkably also in spontaneously occurring large canine tumors (unpublished data). Cullinan Amber is developing CLN-617, a fusion protein that uniquely combines IL-2 and IL-12 with albumin and a collagen-binding domain in a single polypeptide. CLN-617 aims to increase cytokine persistence and retention in the TME upon IT administration and thus improve tolerability. Data presented in this abstract is generated using a murine surrogate, mCLN-617. mCLN-617 retains bioactivity of both IL-2 and IL-12 in the presence of collagen binding. Upon IT injection into B16F10 tumor-bearing mice, systemic drug levels of mCLN-617 were &amp;lt 5% compared to mice dosed via intravenous (IV) administration. When IL-2/IL-12 fusion proteins lacking a collagen-binding domain were injected IT in B16F10-bearing mice, 60% of mice were euthanized due to severe body weight loss, whereas mCLN-617-treated mice maintained their body weight. In checkpoint-refractory B16F10 and MC38 models, mCLN-617 demonstrated 95% tumor growth inhibition and 100% CRs, respectively. All mice cured of their primary MC38 tumors were either protected from re-challenge or showed delayed tumor growth kinetics. Notably, 70% CRs were observed in the MC38 model after a single-dose treatment of mCLN-617. In mice bearing dual-flank MC38 tumors, only one of which was treated IT, 90% of treated tumors and 100% of distal untreated tumors were eliminated when mCLN-617 was combined with an anti-PD1 antibody, demonstrating a curative abscopal response. Of note, mCLN-617 monotherapy in the treated tumor also significantly delayed the growth of distal untreated tumors. Our human lead candidate CLN-617 is produced at high titers in CHO cells and is currently in IND enabling studies. We have demonstrated that the use of collagen-binding domains for tumor retention enables safe and effective delivery of a combination of IL-2 and IL-12 in a single multifunctional molecule. The strong anti-tumor efficacy of mCLN-617 as a monotherapy and significant deepening of responses in combination with checkpoint blockade, together with a favorable tolerability profile, supports future clinical development of CLN-617. Citation Format: Naveen K. Mehta, Bochong Li, Kavya Rakhra, K Dane Wittrup, Patrick A. Baeuerle, Jennifer S. Michaelson. CLN-617 is an IL-2/IL-12 fusion protein with a collagen-anchoring domain that induces potent systemic anti-tumor immunity upon intra-tumoral administration [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 3505.

Differential and organ-specific functions of organic solute transporter α and β in experimental cholestasis

Background & AimsOrganic solute transporter (OST) subunits OSTα and OSTβ facilitate bile acid efflux from the enterocyte into the portal circulation. Patients with deficiency of OSTα or OSTβ display considerable variation in the level of bile acid malabsorption, chronic diarrhea, and signs of cholestasis. Herein, we generated and characterized a mouse model of OSTβ deficiency.MethodsOstβ-/- mice were generated using CRISR/Cas9 and compared to wild-type and Ostα-/- mice. OSTβ was re-expressed in livers of Ostβ-/- mice using adeno-associated virus serotype 8 vectors. Cholestasis was induced in both models by bile duct ligation (BDL) or 3.5-diethoxycarbonyl-1.4-dihydrocollidine (DDC) feeding.ResultsSimilar to Ostα-/- mice, Ostβ-/- mice exhibited elongated small intestines with blunted villi and increased crypt depth. Increased expression levels of ileal Fgf15, and decreased Asbt expression in Ostβ-/- mice indicate the accumulation of bile acids in the enterocyte. In contrast to Ostα-/- mice, induction of cholestasis in Ostβ-/- mice by BDL or DDC diet led to lower survival rates and severe body weight loss, but an improved liver phenotype. Restoration of hepatic Ostβ expression via adeno-associated virus-mediated overexpression did not rescue the phenotype of Ostβ-/- mice.ConclusionsOSTβ is pivotal for bile acid transport in the ileum and its deficiency leads to an intestinal phenotype similar to Ostα-/- mice, but it exerts distinct effects on survival and the liver phenotype, independent of its expression in the liver. Our findings provide insights into the variable clinical presentation of patients with OSTα and OSTβ deficiencies.Lay summaryOrganic solute transporter (OST) subunits OSTα and OSTβ together facilitate the efflux of conjugated bile acids into the portal circulation. Ostα knockout mice have longer and thicker small intestines and are largely protected against experimental cholestatic liver injury. Herein, we generated and characterized Ostβ knockout mice for the first time. Ostα and Ostβ knockout mice shared a similar phenotype under normal conditions. However, in cholestasis, Ostβ knockout mice had a worsened overall phenotype which indicates a separate and specific role of OSTβ, possibly as an interacting partner of other intestinal proteins.

Absent in Melanoma 2 (AIM2) Regulates the Stability of Regulatory T Cells.

Absent in melanoma 2 (AIM2) is a cytosolic dsDNA sensor that has been broadly studied for its role in inflammasome assembly. However, little is known about the function of AIM2 in adaptive immune cells. The purpose of this study was to investigate whether AIM2 has a cell-intrinsic role in CD4+ T cell differentiation or function. We found that AIM2 is expressed in both human and mouse CD4+ T cells and that its expression is affected by T cell receptor (TCR) activation. Naïve CD4+ T cells from AIM2-deficient (Aim2−/−) mice showed higher ability to maintain forkhead box P3 (FOXP3) expression in vitro, while their capacity to differentiate into T helper (Th)1, Th2 or Th17 cells remained unaltered. Transcriptional profiling by RNA sequencing showed that AIM2 might affect regulatory T cell (Treg) stability not by controlling the expression of Treg signature genes, but through the regulation of the cell’s metabolism. In addition, in a T cell transfer model of colitis, Aim2−/−-naïve T cells induced less severe body weight loss and displayed a higher ability to differentiate into FOXP3+ cells in vivo. In conclusion, we show that AIM2 function is not confined to innate immune cells but is also important in CD4+ T cells. Our data identify AIM2 as a regulator of FOXP3+ Treg cell differentiation and as a potential intervention target for restoring T cell homeostasis.

Pathogenesis and genetic characteristics of low pathogenic avian influenza H10 viruses isolated from migratory birds in South Korea during 2010-2019.

Human infection by avian-origin subtype H10 influenza viruses has raised concerns about the pandemic potential of these microbes. H10 subtype low pathogenic avian influenza viruses (LPAIVs) have been isolated from wild birds and poultry worldwide. Here, we isolated 36 H10 LPAIVs from wild bird habitats (a mean annual rate of 3.8% of all avian influenza virus isolations) from January 2010 to April 2019 through a nationwide active surveillance program for avian influenza viruses (AIVs). Phylogenetic analysis revealed that the haemagglutinin (HA) gene of H10 isolates formed eight distinct genetic subgroups (HA-A-H). Unlike other Eurasian-origin subgroups, the HA-H subgroup belonged to the North American lineage. Gene-constellation analysis revealed that 24 H10 LPAIVs constituted ≥18 distinct genotypes, representing high levels of genetic diversity. An intravenous pathogenicity index (IVPI) experiment showed that the pathogenicity of representative strains of the HA-B, E and G subgroups possessing an IVPI score>1.2 was associated with replication capacity in the chicken kidney in the absence of trypsin. Intranasal inoculation experiments showed that a representative strain of the HA-D subgroup replicated and transmitted in chickens without clinical signs. Subclinical virus shedding in chickens may contribute to its silent spread among the poultry population. Moreover, six representative viruses replicated in the lungs of mice without prior adaptation and a representative strain of the HA-C subgroup caused 40% mortality, with severe body weight loss. These findings highlight the importance of intensive surveillance of wild bird habitats, poultry farms and the animal-human interface, along with appropriate risk assessment of isolated viruses.

Co-infection of SARS-CoV-2 and influenza virus causes more severe and prolonged pneumonia in hamsters

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is currently a serious public health concern worldwide. Notably, co-infection with other pathogens may worsen the severity of COVID-19 symptoms and increase fatality. Here, we show that co-infection with influenza A virus (IAV) causes more severe body weight loss and more severe and prolonged pneumonia in SARS-CoV-2-infected hamsters. Each virus can efficiently spread in the lungs without interference by the other. However, in immunohistochemical analyses, SARS-CoV-2 and IAV were not detected at the same sites in the respiratory organs of co-infected hamsters, suggesting that either the two viruses may have different cell tropisms in vivo or each virus may inhibit the infection and/or growth of the other within a cell or adjacent areas in the organs. Furthermore, a significant increase in IL-6 was detected in the sera of hamsters co-infected with SARS-CoV-2 and IAV at 7 and 10 days post-infection, suggesting that IL-6 may be involved in the increased severity of pneumonia. Our results strongly suggest that IAV co-infection with SARS-CoV-2 can have serious health risks and increased caution should be applied in such cases.

1399P Risk factors for body weight loss after gastrectomy for gastric cancer analysed from the JCOG1001 phase III trial

The standard therapy for locally advanced gastric cancer (GC) is a combination of radical gastrectomy and pre- and/or post-operative chemotherapy. Gastrectomy sometimes induces severe body weight loss (BWL) which has been reported as a risk factor for incompliance with an adjuvant chemotherapy. However, there are no reports describing the risk factors for postoperative BWL using data from a prospective clinical trial. The phase III trial (JCOG1001), comparing bursectomy and non-bursectomy in resectable advanced GC treatment, registered a total of 1204 patients between June 2010 and March 2015.