Combinatorial Therapeutic Strategies Research Articles

Effect of probiotics combined with immune checkpoint suppressors and chemotherapeutic agents on digestive system function, intestinal immunity and prognosis in patients with metastatic colorectal carcinoma: a quasi-experimental study

ObjectiveAn exploration of the influence of probiotics combined with immune checkpoint suppressors and chemotherapeutic agents on digestive system function, intestinal immunity and prognosis in patients with metastatic colorectal carcinoma.MethodsThis was a quasi-experimental study. During March 2019 to March 2020, 96 patients with metastatic colorectal carcinoma were arbitrarily classified into control group (n = 48) and intervention group (n = 48). The control group received only immune checkpoint inhibitors or chemotherapy, while the intervention group received immune checkpoint inhibitors or chemotherapy, plus probiotic therapy. Survival and complication rates after 6 months, 12 months and 2 years of treatment were calculated. Intestinal barrier function, immune function and quality of life were analyzed before and after treatment.ResultsNinety-six patients were recorded at the follow-up demonstrating superior survival in the intervention group after 6 months, 12 months and 2 years of therapy. D-lactate and diamine oxidase (DAO) levels were elevated in the intervention group after treatment, with smaller elevations (P < 0.05). The levels of Bifidobacterium and Lactobacillus were remarkably higher in the intervention group after treatment compared to the control group prior to and following therapy (P < 0.05). The amount of Enterococcus and Escherichia coli in the intervention group after treatment was obviously lower compared to the pre-treatment and control groups (P < 0.05). CD3+, CD4+, and CD4+/CD8+ levels were found to be higher in the intervention group (P < 0.05), while CD8+ levels were reduced in the intervention group (P < 0.05). All dimensions of the QLQC-30 scale were higher in the intervention group (P < 0.05). The incidence of complications such as nausea and vomiting, loss of appetite, bloating and diarrhea was considerably reduced in the intervention group (P < 0.05).ConclusionImmune checkpoint suppressors combined with chemotherapeutic agents or probiotic regimens can obviously enhance the prognosis of patients with metastatic colorectal carcinoma. This combination therapeutic strategy can balance the imbalanced intestinal flora, improve intestinal function, and lessen the incidence of adverse events, so it is worthy of clinical application and can lessen the incidence of adverse reactions.

Abstract P018: Mapping cellular and microenvironmental determinants of persister populations in pancreatic cancer following neoadjuvant chemoradiation

Abstract Introduction: The molecular pathways involved in the response to radiation therapy (RT) in pancreatic ductal adenocarcinoma (PDAC) remain poorly understood. Attempts to combine RT with immunotherapies in PDAC have yielded inconsistent outcomes, highlighting a critical gap in our understanding of immune dynamics and mechanisms influencing RT response. Our study aims to uncover molecular pathways and cellular subtypes associated with radioresponse. To do so, we developed a novel cell-cell interaction platform, "SpaCCI", to map ligand-receptor (LR) interactions, essential for understanding cellular communication within the TME and its contribution to therapeutic resistance. Methods: Single-cell RNA sequencing (scRNA-seq) was performed on 16 matched pre- and post-treatment PDAC biopsies from a Phase I/II dose-escalation clinical trial of stereotactic body radiation therapy. Additionally, spatial transcriptomics (ST) was conducted on resected PDAC (20 treated with neoadjuvant chemoradiation, 8 untreated). We integrated scRNA-seq and ST datasets using cell type decomposition and developed SpaCCI to enhance LR interaction detection in tissues with complex cell mixtures. Results: ScRNA-seq identified five molecular subpopulations of cancer cells associated to RT response, with PDAC_3 enriched post-RT and marked by genes involved in lipid metabolism and ferroptosis defense. Post-RT depleted subpopulations, such as PDAC_0 and PDAC_prolif, saw upregulation of pathways related to rapid proliferation and oxidative phosphorylation. In a murine model, PDAC_3 was similarly enriched in post-RT tumors, with persister cell lines from these tumors showing overexpression of ferroptosis defense genes and resistance to ferroptosis inducers including Erastin and RSL3. ST analysis also revealed spatial immune changes post-RT, with increased cytotoxic and effector memory T cells balanced by regulatory T cells and M2-like macrophages, indicating immune escape. NK and memory B cells correlated with improved survival post-RT, while plasma IgG cells localized outside the tumor bed, suggesting compartmentalized immune responses. Using SpaCCI, we observed differential enrichment of ligand-receptor interactions in RT-treated tumors, particularly among lipid-associated tumor macrophages, cancer cells, myofibroblast-like cancer-associated fibroblasts, and endothelial cells. Persister PDAC_3 subpopulations showed enriched LR SEMA3 signaling with CAFs, myeloid cells, and endothelial cells consistent with axon guidance pathways. Conclusions: This study presents a molecular and cellular atlas of PDAC response to RT, revealing persister cells populations with adaptations in lipid metabolism and ferroptosis defense that could be targeted to enhance RT efficacy. ST highlighted complex immune dynamics within the TME demonstrating a balance between inflammatory and immunosuppressive populations. Our findings provide a foundation for combinatorial therapeutic strategies and suggest that axon guidance pathways may also play an important role in the response to RT in PDAC. Citation Format: Vincent Bernard, Li-Ting Ku, Tianyu Wang, Kimal Rajapakshe, Dadi Jiang, Galia Jacobson, Jimin Min, Ching-Wei Tzeng, Manoop S. Bhutani, Paola A. Guerrero, Ethan B. Ludmir, Albert C. Koong, Cullen M. Taniguchi, Anirban Maitra, Ziyi Li, Eugene J. Koay.Mapping cellular and microenvironmental determinants of persister populations in pancreatic cancer following neoadjuvant chemoradiation.[abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Translating Targeted Therapies in Combination with Radiotherapy; 2025 Jan 26-29; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(2_Suppl):Abstract nr P018

Modeling of chemo-radiotherapy targeting growing vascular tumors: A continuum-level approach.

The aim of this study is to demonstrate the enhanced efficiency of combined therapeutic strategies for the treatment of growing tumors, based on computational experiments of a continuous-level modeling framework. In particular, the tumor growth is simulated within a host tissue and treated as a multiphase fluid, with each cellular species considered as a distinct fluid phase. Our model integrates the impact of chemical species on tumor dynamics, and we model -through reaction-diffusion equations- the spatio-temporal evolution of oxygen, vascular endothelial growth factor (VEGF) and chemotherapeutic agents. Simulations of a growing tumor exposed to external radiation showcase the rapid impact of radiotherapy on tumor suppression, however this effect diminishes over time. To enhance the therapeutic efficiency of radiotherapy, we investigate the combination of external radiation with the anti-VEGF drug bevacizumab and the cytotoxic drug docetaxel. Our simulations demonstrate that this synergistic approach integrates the immediate effectiveness of radiation therapy with the enduring tumor-suppressive capabilities of chemotherapy.

Efficacy and Immunological Impact of Intrathecal Baclofen Pumps in Managing Spasticity in Multiple Sclerosis in Adult Patients: A Systematic Review

Background/Objectives: Multiple sclerosis (MS) is a chronic autoimmune disease marked by inflammatory demyelination in the central nervous system, leading to debilitating spasticity. Managing spasticity in MS remains a challenge, and intrathecal baclofen (ITB) therapy has emerged as a potential targeted treatment. This systematic review investigated the efficacy of ITB pumps in managing MS-related spasticity and explored their immunomodulatory effects. Methods: This review adhered to PRISMA guidelines and was submitted for registration retrospectively with the Open Science Foundation. A comprehensive literature search was conducted in PubMed, Embase, Scopus, and Web of Science from January 2013 to August 2024. Studies were included if they examined adult MS patients receiving ITB for spasticity, reporting outcomes related to spasticity and quality of life. Risk of bias was assessed using the Joanna Briggs Institute Critical Appraisal Tools, and findings were synthesized narratively. Results: Eight studies (n = 723 participants) met inclusion criteria. ITB was associated with significant reductions in spasticity severity and improvements in quality of life, with reduced reliance on oral antispasticity medications. Immunologically, ITB has demonstrated potential in modulating inflammatory pathways, downregulating pro-inflammatory cytokines, and shifting immune responses toward an anti-inflammatory profile. Common complications included catheter-related issues and infections, with low overall complication rates. Sensitivity analyses indicated robustness in outcomes across higher-quality studies. Conclusions: ITB pumps are effective in controlling spasticity and offer additional immunological benefits for MS patients. Further research should explore ITB’s long-term immunomodulatory effects and its potential in combined therapeutic strategies. The review was not financially supported, and no conflicts of interest were declared by the authors.

Synergistic Anti-tumorigenic Effects of Cabazitaxel and Usnic Acid Combination on Metastatic Castration-Resistant Prostate Cancer Cells.

Prostate cancer (PC) affects millions of men, causing high mortality rates. Despite the treatment approaches, the options for metastatic castration-resistant prostate cancer (mCRPC), a lethal form of advanced PC, are still limited. Cabazitaxel (Cbx) is the last taxane-derived chemotherapeutic approved for Docetaxel- resistant mCRPC patients. However, its effects are limited due to the activation of several pathways. Therefore, new approaches are needed to increase the efficacy of Cbx. Usnic acid (UA) is a natural product with wellknown anti-tumorigenic and synergistic effects with various chemotherapeutics. Although the cytotoxicity of UA and Cbx has been evaluated on mCRPC cells, the anti-tumorigenic effect of UA combination with any taxane has not been investigated yet. Thus, we aimed to evaluate the possible synergistic effect of Cbx+UA in mCRPC cells. Cell viability and apoptosis were analyzed using WST-1 and Annexin-V. Morphological changes were visualized by fluorescent staining. Finally, cell cycle, mitochondrial health, and ROS levels were determined. Based on WST-1 results, 25 μM UA exhibited significant additive and synergistic effects with the use of Cbx. Annexin V and cell cycle results showed that UA significantly enhanced the Cbx efficacy at increasing doses compared to using only Cbx (**p<0.01). Moreover, combined treatment significantly increased ROS levels and mitochondrial membrane depolarization compared with Cbx alone (**p<0.01). Thus, the results suggest that UA increased the anti-tumorigenic effects of Cbx on mCRPC cells by increasing apoptosis, causing an increase in intracellular ROS and disrupting mitochondrial health. Consequently, combining UA and Cbx offers a new combined therapeutic strategy for mCRPC treatment.

Curcumin reverses high-level tigecycline resistance mediated by different mechanisms in Gram-negative bacteria.

Tigecycline is one of the few effective treatments for multidrug-resistant bacteria. However, the recent emergence and spread of high-level tigecycline resistance in Enterobacteriaceae have significantly limited its clinical use. To combat this challenge, combining antibiotics with adjuvants has emerged as a promising strategy. Curcumin, known for its antibacterial properties and ability to enhance antibiotic efficacy, presents a viable option for reversing tigecycline resistance. This study aimed to evaluate the in vitro and in vivo synergistic effects of curcumin and tigecycline in gram-negative bacteria with different tigecycline resistance mechanisms and to elucidate the molecular mechanisms by which curcumin reverses tigecycline resistance. The checkerboard assay was used to evaluate the synergistic effects of tigecycline and curcumin, while the time-killing curves were used to assess their antibacterial activity. The study also examined their impact on biofilm eradication, the development of tigecycline resistance, and the conjugative transfer of tigecycline-resistant plasmids. The molecular mechanisms underlying the combined effect were investigated. Additionally, the in vivo efficacy of the tigecycline-curcumin combination against tigecycline-resistant Escherichia coli was assessed using a mouse peritonitis infection model. This study revealed that curcumin and tigecycline exhibited synergistic effects against tigecycline-resistant gram-negative bacteria with various resistance mechanisms in vitro. Notably, the addition of curcumin delayed the development of tigecycline resistance in E. coli and impeded the horizontal transfer of tet(X4)-positive IncX1 plasmid. The curcumin-tigecycline combination significantly disrupted cell membrane integrity and reduced efflux pump activity by modulating proton dynamics and inhibiting ATP synthesis. Transcriptomic and proteomic analyses supported these findings, revealing disruptions in central carbon metabolism and substantial effects on the electron transport chain. In vivo experiments using a peritonitis model induced by the tet(X4)-carrying E. coli ZZ9DT16R demonstrated that the curcumin-tigecycline combination improved survival rates, reduced bacterial counts, and enhanced liver and spleen histopathology. Furthermore, the expression of the tet(X4) gene was reduced, and molecular docking studies indicated that curcumin binds to diverse tigecycline resistance proteins. This study is the first to reveal the synergistic action of curcumin and tigecycline, highlighting its potential as a combined therapeutic strategy against tigecycline-resistant gram-negative pathogens with various resistance mechanisms.

Nitroxoline evidence amoebicidal activity against Acanthamoeba castellanii through DNA damage and the stress response pathways.

Acanthamoeba castellanii is a widespread unicellular eukaryote found in diverse environments, including tap water, soil, and swimming pools. It is responsible for severe infections, such as Acanthamoeba keratitis and granulomatous amebic encephalitis, particularly in individuals with immunocompromisation. The ability of protozoans to form dormant and persistent cysts complicates treatment, as current therapies are ineffective against cyst stages and suffer from poor specificity and side effects. Nitroxoline, a quinoline derivative with well-established antibacterial, antifungal, and antiviral properties, is a promising therapeutic candidate. This study aimed to elucidate cellular signalling events that counteract the effects of nitroxoline. In this study, nitroxoline significantly reduced the viability of A. castellanii trophozoites in a dose- and time-dependent manner, inducing morphological changes and apoptosis. Transcriptomic analysis revealed substantial alterations in gene expression, including enrichment of metabolic pathways, DNA damage responses, and iron ion binding. Nitroxoline treatment upregulated genes involved in DNA repair and oxidative stress response while regulating genes in the methionine and cysteine cycles. It also decreased the mitochondrial membrane potential, H₂S production, and total iron amount in A. castellanii. Bioinformatic analyses and molecular docking studies suggest direct interactions between nitroxoline and several A. castellanii proteins. Our research provides a comprehensive molecular map of the response of A. castellanii to nitroxoline, revealing significant changes in gene expression related to the stress response and metabolic pathways. These findings underscore the potential of nitroxoline as a potent anti-Acanthamoeba agent, offering new insights into its mechanism of action and paving the way for effective combinational therapeutic strategies.

Anti-Inflammatory 8-Shogaol Mediates Apoptosis by Inducing Oxidative Stress and Sensitizes Radioresistance in Gastric Cancer.

Radiotherapy is a powerful tumor therapeutic strategy for gastric cancer patients. However, radioresistance is a major obstacle to kill cancer cells. Ginger (Zingiber officinale Roscoe) exerts a potential function in various cancers and is a noble combined therapy to overcome radioresistance in gastric cancer radiotherapy. In this study, we suggested that 8-shogaol, a monomethoxybenzene compound extracted from Zingiber officinale Roscoe, has an anti-cancer and anti-inflammatory activity. In lipopolysaccharide (LPS)-induced inflammatory murine models in vivo and in vitro, 8-shogaol suppressed LPS-mediated cytokine production, including COX-2, TNFα, IL-6, and IL-1β. In xenograft mouse models of AGS gastric cancer cell lines, 8-shogaol reduced tumor volume. In gastric cancer cell lines AGS and NCI-N87, 8-shogaol reduced cell viability and increased caspase-3 activity and cytotoxicity LDH. However, combined with Z-VAD-FMK, 8-shogaol blocked caspase-dependent apoptotic cell death. 8-Shogaol induced intracellular reactive oxygen species (ROS) production, intracellular calcium (Ca2+) release, and endoplasmic reticulum (ER) stress response via the PERK-CHOP signaling pathway. Thapsigargin (TG), an ER stressor, mediated synergistic apoptosis and cell death in 8-shogaol-treated AGS and NCI-N87 cell lines. Nevertheless, loss of PERK or CHOP function suppressed ER-stress-induced apoptosis and cell death in 8-shogaol-treated AGS and NCI-N87 cell lines. 8-Shogaol-induced NADPH oxidase 4 (NOX4) activation is related to ROS generation. However, NOX4 knockdown and ROS inhibitors DPI or NAC blocked ER-stress-induced apoptosis by suppressing the inhibition of cell viability and the enhance of caspase-3 activity, intracellular ROS activity, and cytotoxicity LDH in 8-shogaol-treated AGS and NCI-N87 cell lines. Radioresistant gastric cancer models (AGSR and NCI-N87R) were developed and combined with 8-shogaol and radiation (2 Gy) to overcome radioresistance via the upregulation of N-cadherin and vimentin and the downregulation of E-cadherin. Therefore, these results indicated that 8-shogaol is a novel combined therapeutic strategy in gastric cancer radiotherapy.

Biocompatible nanoparticles self-assembled by PEGylated polyphosphoesters for combination of photodynamic therapy and hypoxia-activated chemotherapy against breast cancer

IntroductionAlthough photodynamic therapy (PDT) shows considerable potential for cancer treatment due to its precise spatial control and reduced toxicity, effectively eliminating residual cells under hypoxic conditions remains challenging because of the resistance conferred by these cells.MethodsHerein, we synthesize an amphiphilic PEGylated polyphosphoester and present a nanocarrier (NPCT) specifically designed for the codelivery of hydrophobic photosensitizer (chlorin e6, Ce6) and hypoxia-activated prodrugs (tirapazamine, TPZ). We investigate the antitumor effect of NPCT on both cellular and animal level.ResultsThe efficient encapsulation of Ce6 and TPZ by NPCT enables the prolonged blood circulation and improved tumor distribution of both agents. Upon internalization by tumoral cells, 660 nm laser irradiation activates Ce6, leading to the generation of reactive oxygen species (ROS) that effectively kill murine 4T1 breast cancer cells. Meanwhile, the PDT process consumes a large amount of oxygen to generate the hypoxic microenvironment that activates the liberated TPZ from NPCT. The resulting highly cytotoxic radicals specifically target and induce cytotoxicity in remaining hypoxic cancer cells. Compared to other groups, the combination of NPCT and 660 nm laser irradiation resulted in the most substantial tumor growth inhibition.DiscussionThis innovative approach provides new avenues for the development of advanced delivery systems based on polyphosphoesters and combination therapeutic strategies.

Abstract IA006: Progress, pitfalls and promise: the journey to effective and well tolerated DNA damage response inhibitor combinations in the clinic

A mounting body of evidence now indicates that poly(ADP-ribose) polymerase (PARP) inhibitors have the potential to be used as a foundation for both monotherapy and combination strategies across a wide spectrum of molecular backgrounds and tumor types. There are multiple promising combination therapeutic strategies, such as those with other DNA damage response (DDR) inhibitors such as ATR inhibitors, immune checkpoint inhibitors, and non-DDR-targeted agents that induce "chemical BRCAness". Despite promising efficacy data, several of the combinations have been hampered by supra-additive toxicities, restricting the combination of full monotherapy doses and schedules of component drugs. I will discuss the progress, pitfalls and promise of different classes of DNA damage response (DDR) inhibitor combinations in the clinic, and discuss potential clinical strategies to develop effective and well tolerated combinations to target PARP inhibitor resistance and to widen the application of DDR agents beyond approved indications Citation Format: Timothy A. Progress, pitfalls and promise: the journey to effective and well tolerated DNA damage response inhibitor combinations in the clinic. [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Optimizing Therapeutic Efficacy and Tolerability through Cancer Chemistry; 2024 Dec 9-11; Toronto, Ontario, Canada. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(12_Suppl):Abstract nr IA006

Clinical significance of HER2 in urothelial carcinoma and analysis of its correlation with glycolytic metabolic characteristics.

This study aimed to explore the clinical relevance of Human Epidermal Growth Factor Receptor 2 (HER2) in urothelial carcinoma (UC) and its association with glycolytic metabolic markers, insulin resistance, and beta-cell function, shedding light on potential therapies targeting both HER2 pathways and cancer metabolism. In this retrospective analysis, 237 UC patients from the Departments of Urology and Pathology at Shandong Provincial Hospital were examined. From 1 January 2023, to 1 October 2024, patients underwent HER2 testing using immunohistochemistry (IHC). We investigated the relationships between HER2 expression and metabolic indicators such as the Homeostatic Model Assessment for insulin resistance (HOMA-IR), beta-cell function (HOMA-β), the triglyceride-glucose (TyG) index, and lactate dehydrogenase (LDH) levels. HER2 status was determined using a standardized scoring system from the 2021 Clinical Pathological Expert Consensus on HER2 Testing in UC, China. Statistical analysis followed CDC guidelines, using multivariate logistic regression to assess the independent impacts of HER2 on metabolic traits. Of the 237 evaluated UC samples, 87.76% exhibited positive HER2 expression. A significant correlation was found between positive HER2 status, advanced tumor stages, and increased LDH levels, suggesting a link between HER2 expression and heightened glycolytic activity. No significant relationships were observed between HER2 status and TyG levels, HOMA-IR, or HOMA-B. Subgroup analyses confirmed the consistency of the relationship between HER2 expression and LDH levels across different demographics and lifestyle factors. Our findings confirm the significant role of HER2 as a prognostic marker and therapeutic target in UC. The association of HER2 positivity with advanced tumor stages and high LDH levels underscores its complex involvement in disease progression. This study highlights the need to explore HER2's biological mechanisms further and pursue combined therapeutic strategies.

Updates in novel immunotherapeutic strategies for relapsed/refractory AML.

Acute myeloid leukemia (AML) is a severe hematological malignancy with poor outcomes, particularly in older adults. Traditional treatment options like high-dose chemotherapy often lead to refractory or relapsed AML, with even worse outcomes. New therapies for relapsed and refractory AML are needed, and this review explores the most recent advancements in immunotherapy in AML. Checkpoint Inhibitors utilizing innate or adaptive immune targeting have shown potential to improve AML outcomes when combined with hypomethylating agents and chemotherapy. The use of adoptive cell therapy in AML demonstrates promising early data, however, there is a need for better target selection. Although early in development, both vaccine therapy as well as stimulator of interferon genes (STING) agonists have potential to enhance the innate immune response to overcome AML's immune evasion. Immunotherapy has become a promising approach for AML treatment, especially in refractory and relapsed AML, especially in patients who are not eligible for allogeneic stem cell transplants. Future research should focus on a deeper understanding of the immune microenvironment to identify the most critical targets for optimization, as well as personalized therapeutic combination strategies. Here we present a comprehensive overview of the recent developments in immunotherapy for relapsed and refractory AML.

Elevated MCL1 expression drives esophageal squamous cell carcinoma stemness and induces resistance to radiotherapy.

Esophageal squamous cell carcinoma (ESCC) stands as the sixth most common cause of cancer-related mortality on a global scale, with a strikingly high proportion-over half-of these fatalities occurring within China. The emergence of radiation resistance in ESCC patients significantly diminishes overall survival rates, complicating treatment regimens and reducing clinical outcomes. There is an urgent need to explore the molecular mechanisms that underpin radiation resistance in ESCC, which could lead to the identification of new therapeutic targets aimed at overcoming this resistance. This study aims to elucidate the role of myeloid cell leukemia-1 (MCL1) in ESCC and its association with radioresistance, thereby providing a novel strategy for enhancing the efficacy of radiotherapy. We used The Cancer Genome Atlas (TCGA) database, Genotype-Tissue Expression (GTEx) project and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) of 10 pairs of fresh endoscopic biopsy samples from patients with ESCC to analyze the messenger RNA (mRNA) expression levels of MCL1 in esophageal cancer tissues as compared to normal tissues. Immunohistochemistry (IHC) staining and Western blotting were performed using an anti-MCL1 antibody to visualize protein expression. The mechanism of radioresistance of ESCC was examined by combining bioinformatics analysis, Western blotting, and clone formation and stemness sphere formation assays. The analysis of TCGA database and the results of RT-qPCR indicated that the mRNA level of MCL1 was overexpressed in esophageal cancer tissues. Subsequently, the results of IHC and Western blotting showed that the protein level of MCL1 expression in cancer tissues was significantly higher than that in adjacent normal tissues. Moreover, there was a significant upregulation of MCL1 in ESCC tissues and in radioresistant tissues and cells, with its overexpression correlating with the acquisition of stemness properties in ESCC. In terms of mechanism, MCL1 induced cell cycle arrest by regulating the expression of cyclin D3 and p21 through the JAK-STAT signaling pathway. G0/G1 phase arrest contributed to the stem cell-like phenotype. Blocking JAK-STAT signaling significantly improved the efficacy of radiotherapy for ESCC. These findings indicate that MCL1 is a critical cell cycle regulator that drives the stemness and radioresistance of ESCC and may thus be a potential target in a combined therapeutic strategy aimed at overcoming radioresistance.

A ferroptosis-targeting detachable dissolvable microneedle as a combined therapeutic strategy for radiation-induced oral mucositis

A ferroptosis-targeting detachable dissolvable microneedle as a combined therapeutic strategy for radiation-induced oral mucositis

Integrated analyses of multi-omic data derived from paired primary lung cancer and brain metastasis reveal the metabolic vulnerability as a novel therapeutic target

BackgroundLung cancer brain metastases (LC-BrMs) are frequently associated with dismal mortality rates in patients with lung cancer; however, standard of care therapies for LC-BrMs are still limited in their efficacy. A deep understanding of molecular mechanisms and tumor microenvironment of LC-BrMs will provide us with new insights into developing novel therapeutics for treating patients with LC-BrMs.MethodsHere, we performed integrated analyses of genomic, transcriptomic, proteomic, metabolomic, and single-cell RNA sequencing data which were derived from a total number of 154 patients with paired and unpaired primary lung cancer and LC-BrM, spanning four published and two newly generated patient cohorts on both bulk and single cell levels.ResultsWe uncovered that LC-BrMs exhibited a significantly greater intra-tumor heterogeneity. We also observed that mutations in a subset of genes were almost always shared by both primary lung cancers and LC-BrM lesions, including TTN, TP53, MUC16, LRP1B, RYR2, and EGFR. In addition, the genome-wide landscape of somatic copy number alterations was similar between primary lung cancers and LC-BrM lesions. Nevertheless, several regions of focal amplification were significantly enriched in LC-BrMs, including 5p15.33 and 20q13.33. Intriguingly, integrated analyses of transcriptomic, proteomic, and metabolomic data revealed mitochondrial-specific metabolism was activated but tumor immune microenvironment was suppressed in LC-BrMs. Subsequently, we validated our results by conducting real-time quantitative reverse transcription PCR experiments, immunohistochemistry, and multiplexed immunofluorescence staining of patients’ paired tumor specimens. Therapeutically, targeting oxidative phosphorylation with gamitrinib in patient-derived organoids of LC-BrMs induced apoptosis and inhibited cell proliferation. The combination of gamitrinib plus anti-PD-1 immunotherapy significantly improved survival of mice bearing LC-BrMs. Patients with a higher expression of mitochondrial metabolism genes but a lower expression of immune genes in their LC-BrM lesions tended to have a worse survival outcome.ConclusionsIn conclusion, our findings not only provide comprehensive and integrated perspectives of molecular underpinnings of LC-BrMs but also contribute to the development of a potential, rationale-based combinatorial therapeutic strategy with the goal of translating it into clinical trials for patients with LC-BrMs.

Supramolecular Assembly‐Enabled Transdermal Therapy of Hypertrophic Scarring Through Concurrent Ferroptosis‐Apoptosis

AbstractHypertrophic scar (HS) remains a major challenge for clinicians due to unsatisfactory therapeutic performance. Although inducing apoptosis of HS fibroblasts (HSFs) has proved to be an effective nonsurgical treatment strategy, potential chemotherapy resistance to apoptosis of HSFs makes the development of new and efficient strategies highly demanding. Herein, a ferroptosis‐apoptosis combined therapeutic strategy for the treatment of HS is developed through supramolecular self‐assembly between cucurbit[7]uril (CB[7]) and two bioactive agents, dihydroartemisinin (DHA) and gold nanoclusters (AuNCs). The resulting self‐assembled supramolecular nanoparticles, named CAD NPs, showed high guest loading efficiency and prominent pH‐responsive degradability in the acidic lysosomes of HSFs. Importantly, both DHA and AuNCs act synergistically to generate excessive reactive oxygen species and lipid peroxidation, leading to mitochondrial damage, and ultimately inducing concurrent ferroptosis and apoptosis on HSFs. Upon further loading into hydrogel microneedles to facilitate their transdermal delivery, these CAD NPs showed superior antiscar therapeutic effects in shortening the treatment span to 3 weeks and improving the HS appearance, as demonstrated at a rabbit ear model of HS. The present supramolecular assembly‐based ferroptosis‐apoptosis strategy provides an innovative guideline for efficiently treating HS as well as other diseases.

Epigallocatechin-3-gallate inhibit the protein arginine methyltransferase 5 and enhancer of Zeste homolog 2 in breast cancer both in vitro and in vivo

PurposeHistone methyltransferases are enzymes that selectively methylate lysine or arginine residues on both histone and non-histone proteins, categorized into lysine methyltransferases and arginine methyltransferases. Notably, EZH2 and PRMT5 are known for catalyzing trimethylation of H3 at K27 and symmetric dimethylation of H4 at R3, respectively. These methylation events are recognized as characteristic histone-repressive marks in cancer. The over expression of PRMT5 and EZH2 were reported in various cancers and recognized as a drug target. The study aims to explore the inhibitory potential of phytocompound, Epigallocatechin-3-gallate (EGCG), against PRMT5 and EZH2 in the breast cancer model. MethodsScreening of an array of phytocompounds was conducted through a combination of in-silico and in-vitro assays. Interactions between EGCG and human PRMT5: MEP50 and EZH2 were evaluated using molecular docking. Binding efficiency was validated, by Surface Plasmon Resonance studies and inhibitory potential was accessed by in vitro methylation followed by western blots, ELISA, and cell-based assays. In-vivo efficacy of EGCG was carried on cell line derived mice xenograft model. ResultsEGCG demonstrated robust interactions with PRMT5:MEP50 complex and EZH2, particularly within the SAM binding site. Surface Plasmon Resonance analysis revealed strong binding affinity in nanomolar concentrations, particularly with PRMT5-MEP50 compared to EZH2. In-vitro assays confirmed EGCG's ability to inhibit PRMT5 and EZH2, leading to a decrease in their catalytic products, namely H4R3me2s and H3K27me3, respectively. EGCG treatment induced both autophagy and apoptosis invitro. In-vivo studies demonstrated significant reductions in tumor size and the proliferation marker ki67, accompanied by a decrease in histone repressive marks. ConclusionThe findings suggest that EGCG effectively inhibits PRMT5 and EZH2, underscoring its potential for combined therapeutic strategies in cancer treatment.

The Role of Immunotherapy in MMR-Deficient Endometrial Carcinoma: State of the Art and Future Perspectives.

This study provides a comprehensive overview of the role of immunotherapy in the treatment of mismatch repair-deficient (MMRd) endometrial carcinomas. Immunotherapy has emerged as a transformative approach in the treatment of MMRd due to the high mutation rate and subsequent PD-1/PD-L1 overexpression seen in these tumors. This review analyzes the current landscape of existing randomized clinical trials, highlighting the efficacy of immune checkpoint inhibitors (ICIs) like pembrolizumab, avelumab, and dostarlimab. Additionally, the focus extends to the potential of combined therapeutic strategies, such as the integration of ICIs with targeted agents, while also exploring the application of immunotherapy in non-traditional settings beyond advanced or recurrent disease. This includes emerging roles in the adjuvant and neoadjuvant contexts to prevent recurrence and target early-stage disease. These findings underscore the importance of tailoring treatments based on the molecular characteristics of each tumor and paving the way for future advancements in the field of gynecologic oncology. Despite promising results, this article acknowledges the necessity of further research to refine patient selection criteria and explore combination strategies that can overcome resistance mechanisms.

Enhancing Tumor Immunity with IL-12 and PD-1 Blockade: A Strategy for Inducing Robust Central Memory T Cell Responses in Resistant Cancer Model.

Background: Although immune checkpoint inhibitors (ICIs) have demonstrated efficacy in treating advanced cancers, their therapeutic success remains limited for many patients, with initial responders often experiencing resistance and relapse. Interleukin-12 (IL-12) is a powerful cytokine for antitumor immunotherapy, enhancing both lymphocyte recruitment into tumors and immune cell activation. Methods: In this study, we successfully produced mouse interleukin-12 (mIL12) through eukaryotic recombinant expression. In vivo, mIL12 exhibited significant control of tumor immunity in ICI-resistant and aggressive tumor models. Further mechanistic analysis indicated that treatment with mIL12 led to a substantial increase in tumor-infiltrating CD4+ T, CD8+ T, cDC1, and CD103+ cDC1 cells. Results: Our data underscore the potential of a combined therapeutic strategy involving IL-12 with PD-1 and CTLA-4 blockade to elicit a potent antitumor immune response. Notably, the co-administration of mIL12 and PD-1 blockade significantly enhanced the presence of central memory T cells (TCM) within tumors. Conclusions: This study is the first to provide evidence that the combination of mIL12 and PD-1 blockers promotes the generation of TCM, potentially contributing to a robust and durable antitumor effect.

Generation of Functional Neurons from Mesenchymal Stem Cells Using Neural Differentiator and Engineered Peptide Hydrogel: Potential Therapeutic Lead for Traumatic Brain Injury.

Traumatic brain injuries (TBIs) cause multifaceted disruption in the neural network, initiate huge inflammation processes, and form glial scars that result in severe damage to the brain. Thus, the treatment of TBI is a challenging task. To address this challenge, a newer and innovative approach is extremely important to develop a successful therapeutic strategy. Toward this aim, we hereby report an extremely effective therapeutic strategy. This interesting approach showcased the development and validation of a combination therapy comprising a neuro-regenerative protective peptide hydrogel (SLNAP) and a potent neuro-regenerative chemical modulator (NCM). It is noteworthy to mention that this hydrogel formulation has injectable nature, which allows it to be applied at focal injury site of brain. Remarkably, our results reveal excellent transdifferentiation of human umbilical cord-derived mesenchymal stem cells (hMSCs) into functional neuron upon treatment with this combination therapeutic formulation. The functionality of regenerated neurons was thoroughly checked through observation of active signals generated from those neurons in electrophysiology recording using patch clamp. Further, we also observed that this strategy not only successfully converts hMSCs into neuron but also spontaneously formed neural stem cells (NSCs) like neurosphere. This work also showcased that this multidomain self-assembling peptide hydrogel emerges as an attractive soft-biomaterial due to its capability of slow and sustained release of the drug at the injury site upon topical application. This resulted in significant regeneration of functional neuron at the injury site. Fascinatingly, we found that this combination therapeutic strategy is highly effective in in vivo brain injury model establishing that this could be a potential and highly effective therapeutic strategy for TBI.