Gastric cancer (GC), characterized by its high incidence and mortality, poses a great threat to public health worldwide. Although various advanced treatments have been developed for GC, the cure rate remains poor. Docetaxel (DTX), a broad-spectrum antitumor drug, has been widely used for the treatment of GC. However, its use in clinical practice is limited by its low water solubility and severe side effects. Our previous work developed a pH/ROS dual-responsive nanoplatform to deliver DTX (DTX/FA-CA-Oxi-αCD NPs) for the targeted treatment of breast cancer. These nanotherapeutics displayed desirable therapeutic effects for breast cancer without obvious adverse effects. On the basis of the treatment potential of DTX/FA-CA-Oxi-αCD NPs, these nanoparticles (NPs) were used for the targeted treatment of GC. In vitro experiments demonstrated that DTX/FA-CA-Oxi-αCD NPs can be efficiently internalized by HGC-27 cells and deeply penetrate tumor spheroids. Moreover, DTX/FA-CA-Oxi-αCD NPs effectively hindered GC cell migration by approximately 60.1

Introduction:Laparoscopic proximal gastrectomy (LPG) with esophagogastrostomy (EG) and double-tract reconstruction (DTR) for early gastric cancer of the upper stomach is a function-preserving surgery that retains oncological curative capability. However, direct comprehensive comparative analyses on the clinical outcomes of EG and DTR as reconstructive techniques following LPG are lacking. Therefore, this study investigated the short- and long-term clinical outcomes of EG and DTR following LPG. Materials and Methods:A retrospective comparative analysis was conducted based on a dataset compiled from two institutions. LPG cases for gastric cancer meeting eligibility criteria were divided into an EG group and a DTR group, and clinicopathological characteristics, perioperative outcomes, nutritional status, 3-year overall survival (OS), and cancer-specific survival (CSS) were analyzed. Furthermore, univariate and multivariate analyses were performed to identify risk factors for complications following LPG. Result:Among the 1198 patients, 104 received LPG, of whom 49 underwent EG and 41 underwent DTR. The DTR was significantly more frequently selected in patients with advanced cancer, while no significant differences were found in the other preoperative clinical factors. No significant differences were observed in perioperative outcomes, including postoperative complications, nutritional parameters, 3-year OS, and CSS. Multivariate analysis identified cardiac disease as a risk factor for postoperative complications following LPG. Conclusion:Short- and long-term clinical outcomes following LPG are equivalent between EG and DTR. For gastric cancer patients with cardiac disease undergoing LPG, careful attention is required for perioperative management.

BACKGROUND:Gastrointestinal (GI) dysmotility is a fundamental clinical issue in multiple diseases such as IBS, and there is a critical requirement for drugs to precisely modulate excessive GI motility. Enteric glial cells (EGCs) are one of the major components of enteric nervous system, which is the key regulator of GI motility. EGCs undergo gliosis in response to multiple stimulation. Among them, mechanical stimulation is the most common stimulation and its motility-promoting effect been proved. However, whether mechanical stimulation could exert an motility-inhibitory effect remained to be elaborated. METHODS:GI motility was assessed by transit time, excreted/retained feces and water content. qPCR, western blotting and immunofluorescence were employed to analyze TRPV4 expression in EGCs. Furthermore, Enteric Gliosis was analyzed by proliferation, activation and neuroinflammation of EGCs. Intracellular Ca2+ concentration were analyzed by Ca2+ imaging. High-throughput sequencing was used to explore the mechanism of TRPV4. RESULTS:Firstly, TRPV4 activation suppressed GI motility in vivo. Secondly, TRPV4 was expressed in EGCs and activation of TRPV4 inhibited GI motility by promoting Enteric Gliosis. Furthermore, TRPV4 activation promotes EGCs Ca2+ signaling, and TRPV4 is required for the regulation of EGCs Ca2+ signaling and gliosis by CaCl2, CaSR, and ATP. Lastly, TRPV4 activation promotes Enteric Gliosis and corrects abnormal GI motility of pathological diarrhea in vivo. CONCLUSION:TRPV4 were identified as a novel accelerator of Enteric Gliosis and suppressor of GI motility. Activation of TRPV4 effectively restoring GI motility homeostasis and offering a potential drug target for gastrointestinal dysmotility.

e14574 Background: TIGIT plays a crucial role in immune regulation, particularly in tumor environments, by acting as an inhibitory receptor. Leveraging its interaction with ligands like CD155 on tumor cells, we developed an innovative CAR-T cell therapy using a mutated TIGIT co-receptor to overcome inhibitory effect induced by CD155, together with targeting Prostate Stem Cell Antigen (PSCA) CAR, chosen for its high expression in bladder and other cancers versus its minimal presence in normal tissues. Methods: We initiated our approach by inducing strategic mutations in TIGIT, selecting a variant with heightened affinity for CD155. Bio-Layer Interferometry (BLI) analysis confirmed the variant's high binding affinity (KD = 1.635 nM). This mutated TIGIT, combined with CD28's signaling components, was integrated as a co-receptor in our CAR construct. For PSCA targeting, we isolated a high-affinity ScFv against PSCA from a fully human phage display library. The specificity of this ScFv for PSCA was rigorously validated using Cell Membrane Protein Targeting Catcher (CMPTC) technology, covering over 6000 human cell membrane proteins. Results: In our comparative analyses, CAR-T cells featuring a mutated TIGIT co-receptor (T-PSCA-CAR-T) matched the cytotoxicity of traditional second-generation BBz-structured CAR-T cells (PSCA-CAR-T) against PSCA-expressing HT-1376 bladder cancer cells. However, T-PSCA-CAR-T cells distinguished themselves by producing significantly higher IFN-γ levels (3805.683±65.506 vs 1271.712±39.768 pg/mL). Our in vivo tumor model, utilizing luciferase-expressing HT-1376 cells, demonstrated T-PSCA-CAR-T's enhanced anti-tumor efficacy, as evidenced by a more significant reduction in bioluminescence signal intensity (average decreasing ratio: 0.02 for T-PSCA-CAR-T vs 0.35 for PSCA-CAR-T; p < 0.05). These results underscore the efficacy of the mutated TIGIT co-receptor in improving CAR-T cell therapy. Conclusions: Our study signifies a transformative advancement in CAR-T cell therapy by incorporating a mutated TIGIT co-receptor, markedly improving therapeutic outcomes in bladder cancer treatment. Utilizing PSCA as a model target not only validates the efficacy of this novel approach but also paves the way for its application in a broader range of cancers, representing a significant leap in the field of immunotherapy.

MEF2D expression in colorectal carcinoma, paired adjacent tissue and normal (S1); Associations of MEF2D expression with clinicopathological features in CRC (S2); Primers used in this study (S3); Antibodies used in this study (S4).

Supplementary Table S1. The primers used for real-time PCR analysis Supplementary Table S2. ABPs obtained from Kyoto Encyclopedia of Genes and Genomes (KEGG) database

High MEF2D expression correlates with cancer cells mesenchymal and invasive features.

MEF2D is involved in various microenvironmental factors induced EMT processes.

MEF2D transcriptionally regulates ZEB1.

Description of additional methods and procedures used in the study. Also includes Supplementary References.

MEF2D is essential for tumorigenesis in vivo.

Legends for Supplementary Figures S1-S4.

S1. The GEPIA website (based on TCGA data) revealed that 48 ABPs were significantly up-regulated in GC tissues. S2. H. pylori induces L-plastin expression in GC cells. S3. Direct contact is required for H. pylori induced L-plastin expression in GC cells. S4. The vacA was not involved in the induction of L-plastin. S5. Decreased expression of L-plastin in presence of U0126 during H. pylori infection. S6. L-plastin expression is increased in GC tissues.

IntroductionAlthough capsaicin has long been used as food additive and medication worldwide, its actions on gastrointestinal tract as its most delivery pathway have not been well addressed.ObjectivesIn the present study, we aimed to study GI actions of capsaicin on mesenteric arterioles in normal and colitis mice and to elucidate the underlying mechanisms.MethodsVasorelaxation of human submucosal arterioles and the mesenteric arterioles from wide-type (WT) mice, TRPV1−/− and TRPV4−/− (KO) mice were measured. The expression and function of TRPV channels in endothelial cells were examined by q-PCR, immunostaining, Ca2+ imaging and membrane potential measurements.ResultsCapsaicin dose-dependently induced vasorelaxation of human submucosal arterioles and mouse mesenteric arterioles in vitro and in vivo through endothelium-dependent hyperpolarization (EDH), nitric oxide (NO), and prostacyclin (PGI2). Using TRPV1 and TRPV4 KO mice, we found that capsaicin-induced vasorelaxation was predominately through TRPV4/EDH, but marginally through TRPV1/NO/PGI2. Capsaicin induced hyperpolarization through activation of endothelial TRPV4 channels and intermediate-conductance of Ca2+-activated K+ channels to finally stimulate vasorelaxation. Importantly, capsaicin exerted anti-colitis action by rescuing the impaired ACh-induced vasorelaxation in WT colitis mice but not in TRPV4 KO colitis mice.ConclusionsCapsaicin increases intestinal mucosal blood perfusion to potentially prevent/treat colitis through a novel TRPV4/EDH-dependent vasorelaxation of submucosal arterioles in health and colitis. This study further supports our previous notion that TRPV4/EDH in mesenteric circulation plays a critical role in the pathogenesis of colitis.

Objective: Robotic surgery has been increasingly used worldwide owing to its advanced features. However, the significant benefits of robotic total gastrectomy (RTG) over laparoscopic total gastrectomy (LTG) have yet to be demonstrated. We conducted a prospective cohort study to compare the safety and efficacy of robotic and laparoscopic total gastrectomy (LTG) with D2 lymphadenectomy for AGC. Methods: Between March 26, 2018 and July 30, 2021, 155 patients between 18 and 80 years of age with locally advanced gastric cancer (cT2-4a, N0/+, MO) were enrolled. The perioperative outcomes within 30 days after surgery were compared between the RTG (n = 69) and LTG (n = 73) groups on a per-protocol (PP) basis. Postoperative complications were evaluated according to the Clavien-Dindo classification. Results: The overall postoperative morbidity rate was 21.74% in the RTG group and 28.77% in the LTG group with no significant difference (P = 0.44), RTG was associated with a lower incidence of pneumonia (4.35% vs. 15.07%, P = 0.047). No mortality was observed in either group. There was no significant difference in the total operative time (284.48 vs. 271.73 min, P = 0.171), but RTG was associated with a lower estimated volume of blood loss (110 vs. 150 ml, P < 0.001) and more total retrieved lymph nodes (LNs) (41.36 vs 35.1, P = 0.019), more extraperigastric LNs (14.91 vs. 12.19, P = 0.024) and more LNs in the suprapancreatic areas (14.68 vs. 11.82, P = 0.017). The laboratory data (amylase, inflammatory, Albumin and T lymphocyte levels) of the RTG group were better than those of the LTG group. Conclusion: According to the results of this prospective cohort study, for patients with locally advanced gastric cancer, Robotic surgery has advantages over laparoscopic surgery for radical total gastrectomy with D2 lymphadenectomy performed by well-trained doctors.

The latest study shows that gastric cancer (GC) ranked the fifth most common cancer (5.6%) with over 1 million estimated new cases annually and the fourth most common cause of cancer death (7.7%) globally in 2020. Metastasis is the leading cause of GC treatment failure. Therefore, clarifying the regulatory mechanisms for GC metastatic process is necessary. In the current study, we discovered that calreticulin (CALR) was highly expressed in GC tissues and related to lymph node metastasis and patient's terrible prognosis. The introduction of CALR dramatically promoted GC cell migration in vitro and in vivo, while the repression of CALR got the opposite effects. Cell migration is a functional consequence of the epithelial-mesenchymal transition (EMT) and is related to adhesion of cells. Additionally, we observed that CALR inhibition or overexpression regulated the expression of EMT markers (E-cadherin, ZO-1, Snail, N-cadherin, and ZEB1) and cellular adhesive moleculars (Fibronectin, integrin β1and MMP2). Mechanistically, our data indicated that CALR could mediate DNA methylation of E-cadherin promoter by interacting with G9a, a major euchromatin methyltransferase responsible for methylation of histone H3 on lysine 9(H3K9me2) and recruiting G9a to the E-cadherin promoter. Knockdown of G9a in CALR overexpressing models restored E-cadherin expression and blocked the stimulatory effects of CALR on GC cell migration. Taken together, these findings not only reveal critical roles of CALR medicated GC metastasis but also provide novel treatment strategies for GC.

Abstract Rationale Neutrophils constitute massive cellular constituents in inflammatory human gastric cancer (GC) tissues, but their roles in pathogenesis of inflammatory T helper (Th) subsets are still unknown. Methods Flow cytometry analysis and immunohistochemistry were used to analyze the responses and phenotypes of neutrophils in different samples from 51 patients with GC. Kaplan‐Meier plots and Multivariate analysis for the survival of patients were used by log‐rank tests and Cox proportional hazards models. Neutrophils and CD4+ T cells were purified and cultured for ex vivo, in vitro and in vivo regulation and function assays. Results GC patients exhibited increased tumoral neutrophil infiltration with GC progression and poor patient prognosis. Intratumoral neutrophils accumulated in GC tumors via CXCL6/CXCL8‐CXCR1‐mediated chemotaxis, and expressed activated molecule CD54 and co‐signaling molecule B7‐H2. Neutrophils induced by tumors strongly expressed CD54 and B7‐H2 in both dose‐ and time‐dependent manners, and a close correlation was obtained between the expressions of CD54 and B7‐H2 on intratumoral neutrophils. Tumor‐derived tumor necrosis factor‐α (TNF‐α) promoted neutrophil activation and neutrophil B7‐H2 expression through ERK‐NF‐κB pathway, and a significant correlation was found between the levels of TNF‐α and CD54+ or B7‐H2+ neutrophils in tumor tissues. Tumor‐infiltrating and tumor‐conditioned neutrophils effectively induced IL‐17A‐producing Th subset polarization through a B7‐H2‐dependent manner ex vivo and these polarized IL‐17A‐producing Th cells exerted protumorigenic roles by promoting GC tumor cell proliferation via inflammatory molecule IL‐17A in vitro, which promoted the progression of human GC in vivo; these effects could be reversed when IL‐17A is blocked. Moreover, increased B7‐H2+ neutrophils and IL‐17A in tumors were closely related to advanced GC progression and predicted poor patient survival. Conclusion We illuminate novel underlying mechanisms that TNF‐α‐activated neutrophils link B7‐H2 to protumorigenic IL‐17A‐producing Th subset polarization in human GC. Blocking this pathological TNF‐α‐B7‐H2‐IL‐17A pathway may be useful therapeutic strategies for treating GC.

BACKGROUND:Leukemia stem cells (LSCs) are responsible for the initiation, progression, and relapse of acute myeloid leukemia (AML). Therefore, a therapeutic strategy targeting LSCs is a potential approach to eradicate AML. In this study, we aimed to identify LSC-specific surface markers and uncover the underlying mechanism of AML LSCs.METHODS:Microarray gene expression data were used to investigate candidate AML-LSC-specific markers. CD9 expression in AML cell lines, patients with AML, and normal donors was evaluated by flow cytometry (FC). The biological characteristics of CD9-positive (CD9+) cells were analyzed by in vitro proliferation, chemotherapeutic drug resistance, migration, and in vivo xenotransplantation assays. The molecular mechanism involved in CD9+ cell function was investigated by gene expression profiling. The effects of alpha-2-macroglobulin (A2M) on CD9+ cells were analyzed with regard to proliferation, drug resistance, and migration.RESULTS:CD9, a cell surface protein, was specifically expressed on AML LSCs but barely detected on normal hematopoietic stem cells (HSCs). CD9+ cells exhibit more resistance to chemotherapy drugs and higher migration potential than do CD9-negative (CD9-) cells. More importantly, CD9+ cells possess the ability to reconstitute human AML in immunocompromised mice and promote leukemia growth, suggesting that CD9+ cells define the LSC population. Furthermore, we identified that A2M plays a crucial role in maintaining CD9+ LSC stemness. Knockdown of A2M impairs drug resistance and migration of CD9+ cells.CONCLUSION:Our findings suggest that CD9 is a new biomarker of AML LSCs and is a promising therapeutic target.

Background The robotic surgical system has several technical advantages over laparoscopic instruments. The technical feasibility and safety of robotic gastrectomy (RG) for gastric cancer have been reported by increasing number of studies. However, the long-term survival and recurrence outcomes after RG for locally advanced gastric cancer (AGC) have seldom been reported. This study aimed to compare long-term oncologic outcomes for patients with locally AGC after RG or laparoscopic gastrectomy (LG). Methods This study comprised 1170 patients underwent RG or LG, respectively, for locally AGC between March 2010 and February 2017. The primary outcome was the 3-year disease-free survival (DFS). The secondary endpoint included 3-year overall survival (OS) and recurrence patterns. One-to-one propensity score matching (PSM) was performed to reduce confounding bias. The outcomes were compared in PSM cohort. Results After PSM, a well-balanced cohort of 816 patients (408 in each group) were included in the analysis. The 3-year DFS rate was 76.2% in the robotic group and 70.1% in the laparoscopic group (P = 0.076). The 3-year OS rates was 76.7% in the robotic group and 73.3% in the laparoscopic group (P = 0.246). In the subgroup analyses for potential confounding variables, neither 3-year DFS nor 3-year OS survival were significantly different between the two groups (all P > 0.05). The two groups showed similar recurrence patterns within 3 years after surgery (P > 0.05). Conclusion For patients with locally AGC, RG can result in comparable long-term survival outcomes without an increase in recurrence rate.

Abstract Actin cytoskeleton dynamic rearrangement is required for tumor cell metastasis and is a key characteristic of Helicobacter pylori (H. pylori)-infected host cells. Actin cytoskeleton modulation is coordinated by multiple actin-binding proteins (ABP). Through Kyoto encyclopedia of gene and genomes database, GEPIA website, and real-time PCR data, we found that H. pylori infection significantly induced L-plastin, a key ABP, in gastric cancer cells. We further explored the regulation and function of L-plastin in H. pylori–associated gastric cancer and found that, mechanistically, H. pylori infection induced gastric cancer cells to express L-plastin via cagA-activated ERK signaling pathway to mediate SP1 binding to L-plastin promoter. Moreover, this increased L-plastin promoted gastric cancer cell proliferation and migration in vitro and facilitated the growth and metastasis of gastric cancer in vivo. Finally, we detected the expression pattern of L-plastin in gastric cancer tissues, and found that L-plastin was increased in gastric cancer tissues and that this increase of L-plastin positively correlated with cagA+ H. pylori infection status. Overall, our results elucidate a novel mechanism of L-plastin expression induced by H. pylori, and a new function of L-plastin–facilitated growth and metastasis of gastric cancer, and thereby implicating L-plastin as a potential therapeutic target against gastric cancer. Implications: Our results elucidate a novel mechanism of L-plastin expression induced by H. pylori in gastric cancer, and a new function of L-plastin–facilitated gastric cancer growth and metastasis, implicating L-plastin as a potential therapeutic target against gastric cancer.