Key Regulator Research Articles

Abstract PR009: FMRP upregulation in cancer: Implicating FMRP-expressing cancer-associated fibroblasts in immune evasion

Abstract Our research has uncovered a novel and significant role for Fragile-X-mental-retardation-protein (FMRP) in cancer biology, extending its well-established functions beyond neuronal development. In a recent Science publication (Zeng et al., 2022), we demonstrated that elevated FMRP levels drive immune evasion in various murine and human tumors by modulating immune cell subsets, particularly through the suppression of effector T-cells. To further elucidate the mechanisms of FMRP-mediated immune evasion, we investigated its influence on the stromal component of the tumor microenvironment. Single-cell RNA sequencing of pancreatic ductal adenocarcinoma (PDAC) tumors revealed cancer-associated fibroblasts (CAFs) as the primary stromal cell subtype expressing FMRP. Given the crucial roles that CAFs play in tumor progression and therapeutic resistance, we explored whether FMRP expression contributes to the immunosuppressive characteristics of CAFs in PDAC and breast tumors. Our findings reveal that FMRP expression is markedly upregulated in CAFs from both breast and PDAC tumors, in contrast to its absence in naïve fibroblasts from healthy tissues. Knockdown of FMRP in CAFs resulted in substantial changes in their myofibroblastic and inflammatory properties, while overexpression of FMRP in naïve fibroblasts was sufficient to induce a CAF-like phenotype. Notably, FMRP overexpression led to significantly elevated inflammatory CAF (iCAF) and myofibroblastic CAF (myCAF) signatures, highlighting its role as a key pan-CAF regulator that influences both extracellular matrix (ECM) modulation and the secretory phenotype of CAFs. We utilized the full-body knockout (KO) mice model of the Fmr1 gene, which encodes FMRP, to further delineate the functional importance of FMRP expression in CAFs in stimulating an immunosuppressive program in vivo. Through bulk RNA sequencing, we developed an FMRP- associated gene signature for CAFs and applied it to human datasets, where we observed a pronounced increase in FMRP signatures in CAFs from human PDAC and breast tumors, highlighting its translational relevance. In conclusion, our research underscores the pivotal role of FMRP in cancer biology, impacting both the immune and stromal components of the tumor microenvironment. Targeting FMRP, particularly within CAFs, emerges as a promising therapeutic strategy to improve cancer treatment outcomes. Citation Format: Simge Yucel, Nadine Fournier, Mireia Blanco Gomez, Douglas Hanahan. FMRP upregulation in cancer: Implicating FMRP-expressing cancer-associated fibroblasts in immune evasion [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr PR009.

Abstract A034: Single cell RNA sequencing of triple negative breast cancer patient-derived xenograft model identifies CRABP1 of cancer-associated fibroblast as a key regulator of breast cancer metastasis

Abstract Triple-negative breast cancer (TNBC) is associated with a high risk of distant metastasis, particularly to the lung and liver. Both intrinsic characteristics of cancer cells and the tumor microenvironment (TME) influence TNBC growth and metastasis. The TME, comprising stromal, immune, and endothelial cells, exhibits heterogeneity both between and within cell types. Identifying the molecular characteristics of TME cells that affect cancer progression is crucial. We aim to investigate this using single-cell RNA sequencing of patient-derived xenograft (PDX) models to explore TME cells and genes involved in TNBC metastasis. We established PDX models by transplanting tumor tissues from 26 TNBC patients into immunodeficient mice. These models were categorized based on their metastatic potential and patient outcomes: non-metastatic (n=5) and metastatic (n=4). Single-cell RNA sequencing of nine PDX tumors revealed significant differences in gene expression of murine stromal cells between metastatic and non-metastatic models. We identified ten genes (Serpinb2, Spp1, Tnc, Thbs1, Timp1, Il11, Mt2, Crabp1, Cck, Mt1) that were highly expressed in the stromal cells of metastatic PDX models. To test if TNBC cells influence these fibroblast genes, we exposed NIH3T3 fibroblasts to conditioned media from the 4T1 TNBC cell line. In three independent experiments, only Crabp1 expression in NIH3T3 cells was consistently increased by 4T1- conditioned media. CRABP1, a retinoic acid-binding protein, is known for its role in differentiation and proliferation by regulating MAPK signaling. Although Crabp1-positive cancer-associated fibroblasts have been noted, its role in TNBC metastasis was previously unexplored. We investigated CRABP1 function using a CRABP1 knockdown NIH3T3 cell line. CRABP1 knockdown led to reduced invasion and migration of 4T1 cells in trans-well assays and decreased invasiveness of 4T1 spheroids in a collagen matrix. Additionally, CRABP1 knockdown NIH3T3 cells showed reduced proliferation in vitro and fewer alpha-SMA positive cancer-associated fibroblasts in co-injected 4T1 tumors in BALB/C mice. In summary, CRABP1, identified through single-cell RNA sequencing of stromal cells in TNBC PDX models, is a potential regulator of TNBC metastasis, affecting cancer cell migration, invasion, and fibroblast proliferation. Citation Format: Woohang Heo, Yujeong Her, Sieun Yang, Dakyung Lee, Rokhyun Kim, Jong-Il Kim, Hyeong-Gon Moon. Single cell RNA sequencing of triple negative breast cancer patient-derived xenograft model identifies CRABP1 of cancer-associated fibroblast as a key regulator of breast cancer metastasis [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A034.

Abstract C027: The role of Liver Receptor Homolog 1 (LRH-1) in regulating cancer progression by modulating the immune response

Abstract Breast cancer is a significant global health concern, being the second most common type of cancer and the second leading cause of cancer-related deaths among women in the U.S. Metastatic breast cancer is responsible for most of these deaths, and current FDA-approved immunotherapies are effective only for a small subset of patients with PD-L1-positive tumors. Thus, there is a critical need for new therapeutic targets to enhance the immune response against breast cancer. Elevated plasma cholesterol concentrations are associated with poor breast cancer outcomes. Our previous studies indicated that myeloid immune cells are highly sensitive to changes in cholesterol levels and other proteins involved in regulating cholesterol homeostasis. However, the role of Liver Receptor Homolog 1 (LRH-1), a key regulator of cholesterol homeostasis, in myeloid cells is not well understood. Notably, higher expression of LRH-1 mRNA in breast tumors is associated with longer time to recurrence and improved overall survival. Therefore, we aimed to investigate the role of LRH-1 in myeloid immune cells and its impact on breast cancer progression. Initial analysis indicated that LRH-1 is expressed in myeloid cells, with particularly high levels observed in neutrophils. Therefore, we examined how LRH-1 regulates neutrophil functions important to cancer biology, including migration, NETosis, and phagocytosis, as well as its influence through neutrophils on T cells. We found that LRH-1 inhibited neutrophil migration toward cancer cells. Since tumor infiltrating neutrophils are generally associated with poor prognosis, LRH-1 reducing neutrophil migration could indicate a decreased ability of these immune-suppressive cells to infiltrate tumors. Importantly, neutrophils treated with an LRH-1 agonist showed reduced NETosis, while antagonist led to increased NETosis. LRH-1 agonist treatment also significantly reduced neutrophil phagocytosis, whereas an antagonist had no notable effect. Furthermore, T cells exhibited increased expansion when co-cultured with neutrophils treated with an LRH-1 agonist, and reduced proliferation with neutrophils pretreated with an LRH-1 antagonist or inverse agonist, particularly in CD4+ helper T cells and later divisions. Since neutrophils were previously implicated in invoking recurrence from dormant lesions, we evaluated LRH-1 in this context. The treatment of mice harboring dormant D2.0R lesions with an LRH-1 inverse agonist resulted in sooner recurrence and metastatic outgrowth compared to vehicle treated mice. In summary, our findings suggest that LRH-1 plays a crucial role in regulating various neutrophil functions, including migration, NETosis, phagocytosis, and T cell expansion. The ultimate consequence of these regulations are control of dormant mammary cancer lesions. These results highlight LRH-1 as a potential therapeutic target for cancer treatment or prevention of metastatic recurrence. Funding: National Cancer Institute (ERN: R01CA234025) Department of Defense (ERN: BCRP Era of Hope Award) Citation Format: Yu Wang, Bryan Duong, Natalia Krawczynska, Shruti V. Bendre, Claire P. Schane, Erin Weisser, Lara Kockaya, Yifan Fei, Anasuya Das Gupta, Hashni E. Vidana Gamage, Adam T. Nelczyk, Dhanya Pradeep, Erik R. Nelson. The role of Liver Receptor Homolog 1 (LRH-1) in regulating cancer progression by modulating the immune response [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C027.

Role of apoptosis-associated proteins p53 and bcl-2 in the pathogenesis of nervous system diseases

Diseases of the central nervous system occupy a leading place, along with cardiovascular and oncological diseases, and the proportion of patients suffering from diseases of the nervous system is increasing as the population ages. This group of diseases includes acute conditions, such as ischemic stroke, and chronic multifactorial diseases — Alzheimer's and Parkinson's diseases, epilepsy, etc. The development of specific methods for their treatment is difficult, and these drugs are not very effective. Almost all brain diseases are based on common mechanisms such as oxidative stress, inflammation and neuronal death. Most often, cells die by apoptosis due to an imbalance between pro-apoptotic and anti-apoptotic factors. This work examines two of them: the apoptosis-promoting transcription factor and tumor suppressor p53 and its opposing B-cell lymphoma protein Bcl-2. The choice of these proteins for study is due to the fact that both proteins are key regulators of apoptosis and are important in the pathogenesis of nervous diseases, since neurons are not highly proliferating cells. The p53 protein is involved in the regulation of many genes responsible for DNA repair, apoptosis, and other biochemical cellular processes; this is especially important when studying neuronal pathology. Bcl-2 suppresses apoptosis in various cells, including neurons, by controlling mitochondrial membrane permeability and inhibiting caspases. In diseases, its expression can either increase, for example, in the case of malignant tumors, or decrease, as in the case of neurodegenerative processes. It has been established that p53 and Bcl-2 are in close interaction in the process of regulating apoptosis; their ratio may be an important prognostic factor. The purpose of this work was to assess the role of these proteins in the pathogenesis of various diseases of the nervous system, and to search for general patterns of changes in their expression and coexpression.

Abstract C021: Tumor-associated microbiota suppresses anti-tumor immunity by driving cDC1 dysfunction in lung cancer

Abstract Type 1 conventional dendritic cells (cDC1s) are critical for initiating and sustaining tumor-reactive CD8 T cells; lack of functional cDC1s is a key driver for failed tumor immunosurveillance and immunotherapy. However, what factors cause the cDC1 insufficiency in the tumor microenvironment (TME) remain poorly understood. Commensal microbiota has emerged as a key regulator of TME and the efficacy of cancer immunotherapies. Nonetheless, most studies have focused on the intestinal microbiome; it remains underexplored whether and how the local microbiota within tumors influences the anti-cancer immunity. Using the autochthonous Kras LSL-G12D/+ ; p53 flox/flox (KP) mouse model of lung cancer, we found an increased local bacterial burden and altered bacterial composition associated with lung tumors. Importantly, this local microbiota suppresses the anti-tumor CD8 T cell response both at baseline and in response to ICB therapies. Specifically, single-cell RNA-seq analysis revealed that the microbiota shapes the functional subsets of tumor-reactive CD8 T cells. Bacterial depletion promotes the accumulation of stem-like Tpex cells (precursor of exhausted T cells) in the TME, which serve as a reservoir of effector CD8 T cells and mediate an effective response upon ICB treatment. To distinguish the function of the local microbiota within TME from the distal gut microbiota, we developed a new method of aerosolized antibiotic treatment, which selectively ablates the intra-tumoral lung microbiota while preserving the intact gut microbiota. Using this approach, we demonstrated that removal of the intra-tumoral lung microbiota markedly improves the tumor-reactive T cell response and sensitizing “cold” lung tumors to ICB therapies. Mechanistically, we identified cDC1s as a central player in microbiota-mediated suppression of anti-tumor immunity. Local microbiota diminishes cDC1 quality and quantity in the lung TME, resulting in impaired T cell priming and Tpex maintenance. At the molecular level, our data revealed that bacterial products derived from the local microbiota induce type I interferon expression in tumor-associated myeloid cells and enhance GM-CSF production from cancer cells: chronic type I interferon and GM-CSF signaling acts cooperatively to reduce the functional cDC1 population. Combining in vitro cDC1 assays, competitive bone marrow chimera experiments and cDC1-specific knockout models, we establish the causal relationships between the local microbiota, IFN-I and GM-CSF signaling in cDC1s, and the tumor response to ICB. Altogether, our study uncovered a dominant role of tumor-associated microbiota in inhibiting cDC1s and suppressing anti-tumor immunity through type I IFN and GM-CSF dependent pathways in lung cancer. Our findings not only identify the intra-tumoral microbiota as a critical therapeutic target for lung cancer treatment, but also provide new insights into the tissue-specific, context-dependent regulatory mechanisms of cDC1s in the TME. Citation Format: Qiang Dong, Chengcheng Jin. Tumor-associated microbiota suppresses anti-tumor immunity by driving cDC1 dysfunction in lung cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C021.

Small, but mitey: investigating the molecular genetic basis formite domatia development and intraspecific variation in Vitisriparia using transcriptomics.

Here, we investigated the molecular genetic basis of mite domatia, structures on the underside of leaves that house mutualistic mites, and intraspecific variation in domatia size in Vitis riparia (riverbank grape). Domatia and leaf traits were measured, and the transcriptomes of mite domatia from two genotypes of V. riparia with distinct domatia sizes were sequenced to investigate the molecular genetic pathways that regulate domatia development and intraspecific variation in domatia traits. Key trichome regulators as well as auxin and jasmonic acid are involved in domatia development. Genes involved in cell wall biosynthesis, biotic interactions, and molecule transport/metabolism are upregulated in domatia, consistent with their role in domatia development and function. This work is one of the first to date that provides insight into the molecular genetic bases of mite domatia. We identified key genetic pathways involved in domatia development and function, and uncovered unexpected pathways that provide an avenue for future investigation. We also found that intraspecific variation in domatia size in V. riparia seems to be driven by differences in overall leaf development between genotypes.

Iron Deficiency Anemia: Etiology, Pathophysiology, Diagnosis, and Treatment Approaches

Iron deficiency anemia (IDA) is a common health problem that affects about 1.24 billion people around the world, mostly children and women of childbearing age. IDA happens when the body doesn't get enough iron, loses too much iron, or can't absorb it properly. This makes erythropoiesis, cellular metabolism, and immune function worse. The World Health Organization says that 40% of women who are pregnant and 32.5% of women who are not pregnant have anemia. People with IDA often feel tired, weak, pale, and have trouble thinking clearly. Lab tests, such as hemoglobin, blood ferritin, and transferrin saturation, are used to make the diagnosis. You can treat the condition in two ways: by taking iron supplements by mouth, such as ferrous sulfate, ferrous gluconate, and ferrous fumarate; or, for serious cases, by giving iron through an IV. Strategies for prevention depend on increasing the amount of iron you get from food, making it more bioavailable, and keeping infections under control. Food addition and fortification programs have been shown to help lower the number of people with IDA. However, problems still exist, especially in areas with poor economies. Recent advance in acknowledging how iron is used and controlled has implications for creating targeted therapeutic approaches. A key regulator of iron balance, hepcidin, is a key player in the pathophysiology of IDA. This review shows how complicated IDA is and how important it is to have treatment plans that consider underlying causes, dietary factors, and socioeconomic factors. Keywords: Iron Deficiency Anemia (IDA), Erythropoiesis, Hemoglobin, Nutritional Deficiencies, Neurological Disorders, Iron Supplements.

Discovery of 1-(Phenylsulfonyl)-1,2,3,4-tetrahydroquinoline Derivative as Orally Bioavailable and Safe RORγt Inverse Agonists for Potential Treatment of Rheumatoid Arthritis.

The retinoic acid receptor-related orphan receptor γt (RORγt) is a key regulator of Th17 cells, associated with autoimmune diseases, making it a significant drug target. Herein, we designed and synthesized 1-(phenylsulfonyl)-1,2,3,4-tetrahydroquinoline derivatives, improving upon GSK2981278 to enhance bioavailability. Of which, D4 exhibited superior bioavailability (F = 48.1 and 32.9% in mice and rats, respectively) compared to GSK2981278 (F = 6.2 and 4.1%, respectively), and effectively treated psoriasis in mice at lower doses. Moreover, for the first time, we report the efficacies of a RORγt inverse agonist in mouse models of rheumatoid arthritis. (R)-D4, the eutomer of D4, matched or exceeded GSK2981278's therapeutic effects at lower doses, with no adverse effects observed after 2 weeks of administration. These results position (R)-D4 as a promising and orally bioavailable candidate for Th17-mediated autoimmune disease treatment.

RESOLVING CONFLICTS OF LAW DURING JUDICIAL PROCEEDINGS: AN EMPIRICAL STUDY OF UKRAINE

Background: Law is a key regulator of social relations. Its systemic nature is fundamental for proper, clear and comprehensive regulation of legal relations. As an integral system, law has its own logic, structure, order and purpose. This purpose is to properly regulate social relations, ensure law and order, and qualitatively and consistently satisfy the rights, obligations and interests of participants in legal relations. However, legal regulation is not without its flaws. One major issue of legal regulation is gaps in the law, where certain social relations lack proper legal regulation due to a lack of specific legislative or legal approaches. Other flaws include conflicts between laws, legislative gaps, qualified silence of the legislator, "darkness" of legal norms, and a "seeming" need for legal regulation and other legal phenomena similar in nature. This article addresses how judges resolve conflicts of law in the course of judicial proceedings, namely the construction of a mechanism to resolve conflicts within national legislation to ensure the right to a fair trial. This issue is of particular importance in the context of the war in Ukraine because, unlike in the relatively stable judicial practice of resolving disputes that arise in a society where there is no war, today, the courts now face unprecedented cases, such as those involving military medical commissions decisions, financial support of military personnel, and new wartime criminal prosecution. Additionally, judges must navigate the procedural norms for the administration of justice in wartime, which are changing rapidly. The study identifies specific cases of conflict in law, particularly in issues related to mobilisation. It highlights how inconsistencies in current legislation and the lack of uniform approaches to overcoming them often prevent citizens from exercising their rights. This situation directly contradicts the Sustainable Development Goals in terms of building peaceful and inclusive societies for sustainable development, ensuring access to justice for all, and building effective, accountable, and inclusive institutions at all levels. The authors of the article highlight the growing importance of legal principles in resolving conflicts within the law. Foundational concepts like the rule of law, legal certainty, and legality are recognised grounds for judicial decision-making. Accordingly, this allows courts to interpret conflicts in specific areas of legislation – such as tax legislation in favour of the taxpayer, child rights in favour of children, and labour law in favour of employees. Consequently, similar claims may be resolved differently depending on their subject matter. To support this analysis, the authors analysed 150 decisions of Ukrainian judges, studied the concept of conflicts, formulated a refined definition, assessed the role of judicial law-making in resolving legal conflicts, and developed a mechanism for addressing conflicts in law by judges. Methods: The authors employed a general dialectical analysis approach grounded in the doctrine of society and thinking, along with the historical method, to analyse the development of legal norms and institutions in different historical periods. This approach provides insights into their origins, evolution and impact on the modern legal system. Methods of analysis and synthesis of information were also utilised. To support the authors’ conclusions, relevant empirical information, including court decisions from the Unified State Register of Court Decisions, were referenced. A total of 150 court decisions containing collisions from 2015 to 2024 were analysed, 22 of which were cited in this article. Results and Conclusions: Conflicts in law are defined as subjectively caused phenomena involving the confrontation of several norms or their totality, resulting in the inability to apply legal norms effectively, clearly, and consistently to regulate social relations. A definition of conflicts in law from a judicial perspective is also proposed: they are contradictions within legal regulation, a negative legal phenomenon that a judge, with the authority vested in them, must resolve in a manner that upholds fundamental principles of law during the administration of justice.

Modulation of SRC by SNTB1 activates the Hippo-YAP pathway during colon adenocarcinoma metastasis.

Colon adenocarcinoma (COAD) ranks as the third most prevalent and lethal cancer in 2020, with metastasis being the primary cause of cancer-related mortality. A comprehensive understanding of the mechanism underlying distant metastasis is imperative for enhancing the prognosis and quality of life of patients with COAD. This study employed gene set enrichment analysis (GSEA) on RNA-sequencing data from 408 patients with COAD in The Cancer Genome Atlas (TCGA) database. GSEA analysis was applied to find the significant hallmark gene set, and genes in the significant hallmark gene set was performed by univariate Cox regression to select the key gene. Then, multivariate Cox regression model was constructed. And various databases were utilized to validate and find the significant oncoprotein and hallmark gene set of the key gene. In addition, the expression of key regulators in para-carcinoma tissue, colon cancer and distant metastases samples were detected by real-time PCR, Immunohistochemistry and Western blot. Additionally, the biological functions were examined by in vitro and in vivo experiments. The scRNA-seq and CellphoneDB were performed to explore cell characters in COAD and the potential mechanism of metastasis. The regulatory network analysis revealed SRC/YAP1 as the most significant oncoprotein and signature gene set associated with SNTB1. Moreover, significant SNTB1 overexpression in COAD metastatic tissues was observed compared to para-carcinoma and primary COAD tissues. Co-immunoprecipitation assays demonstrated the formation of a complex between SNTB1 and SRC proteins. Furthermore, the overexpression of SNTB1 enhanced the proliferation, migration and invasion capacities of COAD cell lines. Caudal vein injection of COAD cells overexpressing SNTB1 in nude mice resulted in increased tumour growth and metastasis to the lung, liver and bone. Finally, single cell RNA-seq revealed alterations in the cellular subtypes of COAD, and CellphoneDB indicated that the interaction between cancer cells exhibiting high SNTB1 expression and enterocytes promoted EMT through cellular communications involving TGF-β, accelerating metastasis in COAD. This study postulates that SNTB1 interacts with SRC to activate the Hippo-YAP pathway, thereby promoting COAD metastasis. Furthermore, cellular communication with enterocytes promotes EMT, facilitating metastasis. These findings propose novel therapeutic targets for preventing or treating metastatic COAD.

Beyond Lux: methods for species and photoreceptor-specific quantification of ambient light for mammals.

Light is a key environmental regulator of physiology and behaviour. Mistimed or insufficient light disrupts circadian rhythms and is associated with impaired health and well-being across mammals. Appropriate lighting is therefore crucial for indoor housed mammals. Light is commonly measured in lux. However, this employs a spectral weighting function for human luminance and is not suitable for 'non-visual' effects of light or use across species. In humans, a photoreceptor-specific (α-opic) metrology system has been proposed as a more appropriate way of measuring light. Here we establish technology to allow this α-opic measurement approach to be readily extended across mammalian species, accounting for differences in photoreceptor types, photopigment spectral sensitivities, and eye anatomy. We develop a high-throughput method to derive spectral sensitivities for recombinantly expressed mammalian opsins and use it to establish the spectral sensitivity of melanopsin from 13 non-human mammals. We further address the need for simple measurement strategies for species-specific α-opic measures by developing an accessible online toolbox for calculating these units and validating an open hardware multichannel light sensor for 'point and click' measurement. We finally demonstrate that species-specific α-opic measurements are superior to photopic lux as predictors of physiological responses to light in mice and allow ecologically relevant comparisons of photosensitivity between species. Our study presents methods for measuring light in species-specific α-opic units that are superior to the existing unit of photopic lux and holds the promise of improvements to the health and welfare of animals, scientific research reproducibility, agricultural productivity, and energy usage.

Neutrophil-centric analysis of gastric cancer: prognostic modeling and molecular insights.

Gastric cancer remains a significant global health concern with poor prognosis. This study investigates the role of neutrophils in gastric cancer progression and their potential as prognostic indicators. Using multi-omics approaches, including Weighted Gene Co-expression Network Analysis (WGCNA), machine learning, and single-cell analysis, we identified neutrophil-associated gene signatures and developed a robust prognostic model. Our findings reveal distinct gastric cancer subtypes based on neutrophil-associated genes, with one subtype showing increased neutrophil infiltration and poorer prognosis. Single-cell analysis uncovered neutrophil-associated alterations in cell composition, gene expression profiles, and intercellular communication within the tumor microenvironment. Additionally, we explored the relationship between neutrophil-associated genes, microbiota composition, and alternative splicing events in gastric cancer. Furthermore, we identified QKI as a key regulator of alternative splicing and demonstrated its role in promoting malignant phenotypes and enhancing TGF-beta signaling and epithelial-mesenchymal transition in gastric cancer cells by wet experiment. Lastly, the role of QKI in the association with drug resistance and the identification of specific agents for treating QKI-associated drug resistance were also explored. This comprehensive study provides novel insights into the complex interplay between neutrophils, the tumor microenvironment, microbiota, alternative splicing and gastric cancer progression, offering potential new targets for therapeutic intervention.

Exploring the Updated Roles of Ferroptosis in Liver Diseases: Mechanisms, Regulators, and Therapeutic Implications.

Ferroptosis, a newly discovered mode of cell death, is a type of iron-dependent regulated cell death characterized by intracellular excessive lipid peroxidation and imbalanced redox. As the liver is susceptible to oxidative damage and the abnormal iron accumulation is a major feature of most liver diseases, studies on ferroptosis in the field of liver diseases are of great interest. Studies show that targeting the key regulators of ferroptosis can effectively alleviate or even reverse the deterioration process of liver diseases. System Xc- and glutathione peroxidase 4 are the main defense regulators of ferroptosis, while acyl-CoA synthetase long chain family member 4 is a key enzyme causing peroxidation in ferroptosis. Generally speaking, ferroptosis should be suppressed in alcoholic liver disease, non-alcoholic fatty liver disease, and drug-induced liver injury, while it should be induced in liver fibrosis and hepatocellular carcinoma. In this review, we summarize the main regulators involved in ferroptosis and then the mechanisms of ferroptosis in different liver diseases. Treatment options of drugs targeting ferroptosis are further concluded. Determining different triggers of ferroptosis can clarify the mechanism of ferroptosis occurs at both physiological and pathological levels.

Regulation and functions of the DLC family of RhoGAP proteins: Implications for development and cancer.

The DLC (Deleted in Liver Cancer) family of RhoGAP (Rho GTPase-activating) proteins has been extensively studied since the identification of the first family member nearly 30 years ago. Rho GTPase signaling is essential for various cellular processes, including cytoskeletal dynamics, cell migration, and proliferation. Members of the DLC family are key regulators of this signaling pathway, with well-established roles in development and carcinogenesis. Here, we provide a comprehensive review of research into DLC regulation and cellular functions over the last three decades. In particular, we summarize control mechanisms of DLC gene expression at both the transcriptional and post-transcriptional level. Additionally, recent advances in understanding the post-translational regulation of DLC proteins that allow for tuning of protein activity and localization are highlighted. This detailed overview will serve as resource for future studies aimed at further elucidating the complex regulatory mechanisms of DLC family proteins and exploring their potential as targets for therapeutic applications.

Astrocytic NHERF-1 Increases Seizure Susceptibility by Inhibiting Surface Expression of TREK-1.

Mature hippocampal astrocytes exhibit a linear current-to-voltage (I-V) K+ membrane conductance called passive conductance. It is estimated to enable astrocytes to keep potassium homeostasis in the brain. We previously reported that the TWIK-1/TREK-1 heterodimeric channels are crucial for astrocytic passive conductance. However, the regulatory mechanism of these channels by other binding proteins remains elusive. Here, we identified Na+/H+ exchange regulator-1 (NHERF-1), a protein highly expressed in astrocytes, as a novel interaction partner for these channels. NHERF-1 endogenously bound to TWIK-1/TREK-1 in hippocampal cultured astrocytes. When NHERF-1 is overexpressed or silenced, surface expression and activity of TWIK-1/TREK-1 heterodimeric channels are inhibited or enhanced, respectively. Furthermore, we confirmed that reduced astrocytic passive conductance by NHERF-1 overexpressing in the hippocampus increases kainic acid (KA)-induced seizure sensitivity. Taken together, these results suggest that NHERF-1 is a key regulator of TWIK-1/TREK-1 heterodimeric channels in astrocytes and suppression of TREK-1 surface expression by NHERF-1 increases KA-induced seizure susceptibility via reduction of astrocytic passive conductance.

DLL4-targeted CAR-T therapy sensitizes neoadjuvant chemotherapy via eliminating cancer stem cells and reshaping immune microenvironment in HER2+ breast cancer

BackgroundNeoadjuvant therapy with trastuzumab, pertuzumab and paclitaxel (THP) has significantly improved the prognosis of patients with human epidermal growth factor receptor 2 (HER2)+ breast cancer (BC). However, there remains a...

Enhancing disease risk gene discovery by integrating transcription factor-linked trans-variants into transcriptome-wide association analyses.

Transcriptome-wide association studies (TWAS) have been successful in identifying disease susceptibility genes by integrating cis-variants predicted gene expression with genome-wide association studies (GWAS) data. However, trans-variants for predicting gene expression remain largely unexplored. Here, we introduce transTF-TWAS, which incorporates transcription factor (TF)-linked trans-variants to enhance model building for TF downstream target genes. Using data from the Genotype-Tissue Expression project, we predict gene expression and alternative splicing and applied these prediction models to large GWAS datasets for breast, prostate, lung cancers and other diseases. We demonstrate that transTF-TWAS outperforms other existing TWAS approaches in both constructing gene expression prediction models and identifying disease-associated genes, as shown by simulations and real data analysis. Our transTF-TWAS approach significantly contributes to the discovery of disease risk genes. Findings from this study shed new light on several genetically driven key TF regulators and their associated TF-gene regulatory networks underlying disease susceptibility.

Enhancing the anticancer effect of metformin through nanoencapsulation: Apoptotic induction, inflammatory reduction, and suppression of cell migration in colorectal cancer cells.

Colorectal cancer (CRC) continues to be a significant health challenge, necessitating the development of efficient therapeutic strategies. Drug repurposing, which involves the use of existing medications for new purposes, presents a promising opportunity. Metformin, a widely used antidiabetic drug, has demonstrated potential anticancer effects. To enhance its efficacy, we formulated nano-metformin, metformin encapsulated within pectin nanoparticles. Our study aimed to evaluate the superiority of nano-metformin over free metformin in treating CRC. The cytotoxicity of both metformin and nano-metformin on Caco-2 CRC cells was assessed using the MTT assay, revealing a significant dose-dependent inhibition of cell growth using nano-metformin. The anti-inflammatory potential was evaluated by measuring the levels of nitric oxide and the pro-inflammatory cytokines IL-2 and IL-6 following lipopolysaccharide (LPS) induction, and the results revealed that treating LPS-induced cells with nano-metformin significantly reduced the production of these inflammatory mediators. To elucidate the mechanism of cell death, we employed an acridine orange/ethidium bromide staining assay, which revealed the enhancement of apoptotic cell death following treatment with nano-metformin. Additionally, we examined the expression of key apoptotic regulators using real-time qPCR. Nano-metformin, in particular, significantly downregulated the expression of the antiapoptotic markers Bcl-2 and Survivin while upregulating the proapoptotic caspases 3, 7, and 9. The comet assay revealed significant DNA damage induced by treatment with the nano-metformin compared with that in the free form. Moreover, nano-metformin significantly reduced the migration ability of cells. In conclusion, our work revealed the superior efficacy of our formulated nanoform over free metformin, highlighting its potential as a promising therapeutic agent for CRC treatment.

Follicular fluid-derived extracellular vesicles miR-34a-5p regulates granulosa cell glycolysis in polycystic ovary syndrome by targeting LDHA.

Polycystic ovary syndrome (PCOS) is highly prevalent in women of reproductive age worldwide, exhibits highly heterogeneous clinical presentation and biochemical parameters, and has no cure. This study aimed to investigate the role of miR-34a-5p in PCOS, its effect on glycolysis in granulosa cells (GCs), and its potential contribution to follicular dysregulation. Herein, Follicular follicular fluid (FF) samples were collected from six patients with PCOS and six healthy controls undergoing in vitro fertilization-embryo transfer. The isolated extracellular vesicles were characterized by transmission electron microscopy, nanoparticle tracking analysis, and western blotting. Additionally, miRNA sequencing was performed to identify differentially expressed microRNAs, and their functions were analyzed through transcriptomics. The In vitro effects of miR-34a-5p on glycolysis, cell proliferation, and apoptosis were assessed in human ovarian granulosa tumour cell line (KGN cells). Targets of miR-34a-5p were identified by dual-luciferase reporter assays, and quantitative real-time polymerase chain reaction and western blotting were performed to determine gene and protein expression. The level of miR-34a-5p in FF-derived extracellular vesicles derived from patients with PCOS was significantly higher than that of the control group. Transcriptomic analysis highlighted miR-34a-5p as a key regulator of glycolysis and apoptosis. Furthermore, in vitro analysis showed that miR-34a-5p targeted lactate dehydrogenase A (LDHA), inhibited glycolysis, reduced energy supply to GCs, and promoted apoptosis of KGN cells. Conversely, miR-34a-5p inhibition restored glycolysis function and cell viability. The findings of this study show that miR-34a-5p mediates GC apoptosis in PCOS by targeting LDHA and inhibiting glycolysis, suggesting its crucial role in PCOS pathophysiology, and offering potential therapeutic targets for improving follicular development and fertility outcomes in patients with PCOS. Further research is needed to explore the clinical implications of miR-34a-5p and its use as a biomarker for early diagnosis and prognosis of PCOS.

Epigenetic characterization of adult rhesus monkey spermatogonial stem cells identifies key regulators of stem cell homeostasis.

Spermatogonial stem cells (SSCs) play crucial roles in the preservation of male fertility. However, successful ex vivo expansion of authentic human SSCs remains elusive due to the inadequate understanding of SSC homeostasis regulation. Using rhesus monkeys (Macaca mulatta) as a representative model, we characterized SSCs and progenitor subsets through single-cell RNA sequencing using a cell-specific network approach. We also profiled chromatin status and major histone modifications (H3K4me1, H3K4me3, H3K27ac, H3K27me3 and H3K9me3), and subsequently mapped promoters and active enhancers in TSPAN33+ putative SSCs. Comparing the epigenetic changes between fresh TSPAN33+ cells and cultured TSPAN33+ cells (resembling progenitors), we identified the regulatory elements with higher activity in SSCs, and the potential transcription factors and signaling pathways implicated in SSC regulation. Specifically, TGF-β signaling is activated in monkey putative SSCs. We provided evidence supporting its role in promoting self-renewal of monkey SSCs in culture. Overall, this study outlines the epigenetic landscapes of monkey SSCs and provides clues for optimization of the culture condition for primate SSCs expansion.