Tumor immunotherapy has demonstrated clinical success in some patients, but its efficacy in most solid tumors is hampered by poor infiltration of tumor-specific cytotoxic T lymphocytes (CTLs). Activating virus-specific bystander T cells via loading viral epitopes onto tumor-associated MHC class I molecules can induce antitumor immunity, but this approach is restricted by MHC polymorphism and individual infection histories. Here, we developed a hepatocellular carcinoma-targeted adeno-associated virus system that directs full-length membrane-anchored expression of the SARS-CoV-2 receptor-binding domain on tumor cells. This design bypasses MHC class I epitope pre-screening by activating pre-existing SARS-CoV-2-specific CTLs to exert potent cytotoxicity, and simultaneously engaging humoral immunity to induce antibody-dependent cell-mediated cytotoxicity. In a murine hepatocellular carcinoma model, our treatment markedly reduced tumor burden. It also enhanced infiltration of pre-existing bystander immune cells, accompanied by increased intratumoral IFN-γ+ CD8+ T cells and perforin-1+ NK cells. These findings demonstrate that leveraging pre-existing SARS-CoV-2 immunity to convert immunologically "cold" tumors into immune-recognizable targets, providing a promising translational strategy for refractory malignancies.

Background and aims Histone lysine lactylation has emerged as a key epigenetic link between metabolism and gene regulation. Its homeostasis depends on both lactylation and delactylation. The biological and clinical relevance of histone lysine delactylation in hepatocellular carcinoma (HCC) remains unclear. This study aimed to investigate the clinical relevance of histone lysine delactylation (HLDL) in hepatocellular carcinoma and to establish an HLDL-based molecular classification for precision therapy. Methods We systematically profiled HLDL across multi-omics HCC cohorts (n > 1,700) to establish an HLDL-based molecular classification. Distinct subtypes were characterized by genomic, transcriptomic, and immune features. Therapeutic susceptibilities were predicted through pharmacogenomic modeling and validated by in vitro modulation of histone lysine lactylation. Results The HLDL classification delineated two major subtypes with distinct biological and therapeutic profiles. Low-HLDL tumors displayed higher genomic instability, activated oncogenic signaling, and an immune-infiltrated yet suppressive microenvironment, corresponding to poorer outcomes but higher sensitivity to immune checkpoint inhibitors, atezolizumab plus bevacizumab therapy, and sorafenib. In contrast, high-HLDL tumors exhibited a metabolically stable and delactylation-active phenotype, responding more favorably to lenvatinib and transarterial chemoembolization. Functional experiments confirmed that reduced HLDL activity led to elevated histone lysine lactylation, PD-L1 upregulation, and lenvatinib resistance, whereas restoring delactylation reversed these effects. Conclusions Histone lysine delactylation constitutes a critical epigenetic determinant of HCC heterogeneity and therapeutic response. The HLDL-based classification developed in this study integrates metabolic, immune, and pharmacologic features into a concise framework, offering new avenues for precision therapy and potential targets for modulating the lactylation-delactylation balance in HCC.

Abstract Efficient intracellular protein delivery represents an essential prerequisite for protein-based biotechnologies and therapeutics targeting intracellular components. However, this process is limited by multiple factors, including nonspecific protein binding, insufficient cellular uptake, inefficient endosomal escape, and inadequate cytosolic protein release. Here we show that by engineering fully recombinant supercharged protein nanocages, we achieve exceptionally high cellular uptake using a strategy we term ‘supercharged interface engineering’. By incorporating unnatural amino acids bearing phenylboronic acid groups, we develop a representative protein nanocage, pFn + . Simply mixing pFn+ with protein cargoes forms a noncovalent complex possessing enhanced cellular uptake efficiency, robust endosomal escape capability, and excellent biocompatibility. Notably, this system successfully delivers functional gene-editing tools and therapeutic antibodies in female mouse models. These findings indicate that pFn+ represents a promising platform for enhancing the cytosolic delivery of protein cargoes. Moreover, the proposed supercharged interface engineering strategy is valuable for advancing next-generation intracellular protein delivery systems.

Microbial therapy has emerged as a promising strategy for cancer treatment, driven by advances in synthetic biology that enable precise engineering of probiotic functions. Engineered bacteria offer several therapeutic advantages, including localized drug delivery, direct tumor killing, and immune modulation. Bacteria-based therapies offer promise for colorectal cancer treatment, yet safety issues of chassis cells and poor targeting remain key challenges. Here, we engineered a novel bacterial system based on the gut symbiont Bacteroides thetaiotaomicron, a naturally abundant and safe resident of the human intestine. By systematically screening BT-derived surface anchors, we established an efficient display system capable of presenting the tumor-targeting protein HlpA on the bacterial surface. Engineered BT demonstrated specific targeting of colorectal tumors in both in vitro cell models and in vivo animal models. Building on this platform, we integrated a therapeutic module encoding hemolysin E, which conferred dual antitumor effects in an AOM/DSS-induced colitis-associated cancer mouse model by alleviating intestinal inflammation and inhibiting tumor growth. Our findings highlight BT as a promising vector for engineered bacterial therapies, providing a new gut symbiont-based targeted cancer treatment strategy.

Microbial therapy has emerged as a promising strategy for cancer treatment, driven by advances in synthetic biology that enable precise engineering of probiotic functions. Engineered bacteria offer several therapeutic advantages, including localized drug delivery, direct tumor killing, and immune modulation. Bacteria-based therapies offer promise for colorectal cancer treatment, yet safety issues of chassis cells and poor targeting remain key challenges. Here, we engineered a novel bacterial system based on the gut symbiont Bacteroides thetaiotaomicron, a naturally abundant and safe resident of the human intestine. By systematically screening BT-derived surface anchors, we established an efficient display system capable of presenting the tumor-targeting protein HlpA on the bacterial surface. Engineered BT demonstrated specific targeting of colorectal tumors in both in vitro cell models and in vivo animal models. Building on this platform, we integrated a therapeutic module encoding hemolysin E, which conferred dual antitumor effects in an AOM/DSS-induced colitis-associated cancer mouse model by alleviating intestinal inflammation and inhibiting tumor growth. Our findings highlight BT as a promising vector for engineered bacterial therapies, providing a new gut symbiont-based targeted cancer treatment strategy.

Glucagon-like peptide-1 receptor (GLP-1R) activation is widely assumed to regulate the metabolic disorder in Alzheimer's disease (AD). However, direct evidence for this hypothesis is lacking, and currently, there is no oral GLP-1R agonist with effective blood-brain barrier-penetrating ability. Here, we show that a candidate peptide, OHP2, an oral GLP-1R agonist with blood-brain barrier permeability, exhibits promising therapeutic potential for AD. OHP2 primarily activates GLP-1R on astrocytes, leading to increased aerobic glycolysis and driving lactate release. Astrocyte-derived lactate is taken up by neurons and elevates histone H3 lysine 9 lactylation (H3K9la), which in turn facilitates lipid transport from neurons back to astrocytes. This astrocyte-neuron metabolic coupling sustains continuous aerobic glycolysis and offers a potential treatment strategy for AD. The H3K9la derived from OHP2 links glucose and lipid metabolic cycle and facilitates metabolic coupling between astrocytes and neurons, which leads to remission of metabolic disturbances in AD. Thus, our study provides a new candidate molecule for drug research in treating AD and illustrates that intracerebral GLP-1R activation, which facilitates astrocyte-neuron metabolic coupling, may be a potential approach for the treatment of AD.

Interleukin-12 (IL-12) is a potent immunostimulatory cytokine that enhances T cell and natural killer (NK) cell activation and promotes Th1-type immune responses. However, its clinical application as monotherapy has been hampered by systemic toxicity and limited efficacy. As a result, IL-12-based combination strategies have attracted increasing attention in cancer therapy. This review summarizes the immune regulatory mechanisms by which IL-12 remodels the tumor microenvironment and enhances antitumor immunity. We discuss recent preclinical and clinical advances in combining IL-12 with immune checkpoint inhibitors, chemotherapy, cellular therapies such as CAR-T cells, and cancer vaccines. In addition, emerging synergistic approaches involving targeted therapies and STING agonists are highlighted. Finally, we address current challenges and future directions, focusing on strategies to improve tumor specificity and minimize systemic toxicity. Overall, this review emphasizes IL-12 as a key immunomodulatory component in next-generation combination cancer therapies.

BACKGROUND:Hepatitis B virus (HBV) is the dominant infectious driver of hepatocellular carcinoma (HCC), but its multilevel effects on the tumor microenvironment (TME) remain incompletely catalogued, thereby constraining the advancement of future research and the refinement of clinical practice. METHODS:A comprehensive search in PubMed was conducted using the keywords "((HBV) or (Hepatitis B virus)) and ((HCC) or (hepatocellular carcinoma))". From a total of 16 473 articles, articles unrelated to tumor microenvironment or therapeutic strategies were rejected. Relevant papers published before June 2025 were included. Priority is given to authoritative articles and the latest research findings. RESULTS:The influence of HBV in the TME of HCC encompasses the generation and development of tumor cells, the number, function, and phenotype of immune cells, as well as the phenotype, characteristics, and role of other cells. Additionally, HBV is involved in reshaping the metabolic microenvironment of HCC and plays a role in the complex network within the microenvironment. The characteristics of the TME in HBV-HCC play a guiding role in the development of novel therapeutic targets, discovery of biomarkers, optimization of risk and stratification models, and improvement of clinical treatment strategies. By fully accounting for tumor heterogeneity and tailoring personalized treatment approaches for different TME subtypes, the precision and efficacy of therapies might be significantly enhanced. CONCLUSION:Deepening the understanding of the TME in HBV-HCC contributes to more precise and effective clinical treatments.

Microglia-mediated neuroinflammation, considered one of the most plausible pathogenic hypotheses underlying Alzheimer’s disease (AD), plays a pivotal role in the initiation and progression of this devastating condition. Recently, glucagon-like peptide-1 receptor agonists (GLP-1RAs) have demonstrated promising neuroprotective effects in both preclinical and clinical studies. Previously, we developed an orally-administered GLP-1RA peptide called OHP2, which is capable of crossing the blood-brain barrier for the treatment of AD. OHP2 has been shown to effectively reduce brain inflammation in AD mouse models. In this study, we discovered that OHP2 treatment induced IL-27 secretion from astrocytes and modulated microglial reprogramming from the neurotoxic M1 phenotype to the neuroprotective M2 phenotype through glycolysis/cGAS lactylation clock/mTOR pathway, thereby alleviating excessive neuroinflammation. These findings provide a rationale for further pharmacological investigations into OHP2 and suggest that IL-27 may hold significant implications for AD therapy as a metabolic regulator.

Engineered therapeutic microbes for intestinal inflammation must be capable of gut colonization, sensitive detection of disease-associated biomarkers and targeted delivery of therapeutic molecules. Examples of microbes displaying all three characteristics are limited. Here we engineered Bacteroides thetaiotaomicron, a human gut commensal bacterium with colonization ability, colonic tropism and innate anti-inflammatory properties, as a chassis to create programmable bacterial strains termed Btbots. We developed genetic circuits that were integrated into the bacterial chromosome to sense two intestinal inflammation biomarkers, deoxycholic acid (DCA) and nitric oxide (NO), and to enhance surface display and secretion of therapeutic molecules in response. Btbots with these biosensors were developed that either display trefoil factor-3 to facilitate mucosal repair or secrete interleukin-35 to suppress inflammation. These Btbots sensed DCA and NO biomarkers and released therapeutic agents, alleviating colitis and modulating the gut microenvironment and microbiota in mouse models. This work establishes a proof of concept for localized sensing and consequent therapeutic molecule release for gastrointestinal applications, whose clinical potential awaits further investigation.

Bacteroides thetaiotaomicron is a common microorganism in the human gut that has been linked to health benefits. Furthermore, it is an emerging synthetic biology chassis with the potential to be modified into diagnostic or therapeutic engineered probiotics. However, the absence of biological components limits its further applications. In this study, we developed an antiterminator microbial whole-cell biosensor (MWCB) based on B. thetaiotaomicron. The antiterminator-based element allows the chassis to detect colitis in mice by responding to nitrate and nitrite in an inflammatory environment. In particular, the nitrate/nitrite-inducible promoter was obtained by combining the constitutive promoter with the inducible terminator. Subsequently, the promoter and RBS were replaced to optimize a sensitive and specific response to nitrate/nitrite. A preliminary in vitro assessment was conducted to ascertain the functionality of the biosensor. Its in vivo sensing ability was evaluated in a chemically induced mouse model of ulcerative colitis (UC). The results demonstrated that the MWCB exhibited a robust response to colitis, with a notable positive correlation between the intensity of the response and the level of inflammation. This novel sensing element may provide a new avenue for the development of components for unconventional chassis, like B. thetaiotaomicron. It will also facilitate the development of engineered probiotics based on B. thetaiotaomicron, thereby providing patients with a wider range of medical treatment options.

Inflammatory bowel disease (IBD) treatment is challenged by limited efficacy and side effects of current therapies. While Astragalus membranaceus polysaccharides (APS) show anti-inflammatory potential, their structure-activity relationship is unclear. This study compared the therapeutic effects of the linear α-1,4-glucan APS-G2 and the branched-chain APS-A1, enzymatically derived APS oligosaccharides. Structure-function analysis revealed that the homogeneous α-1,4-glucan backbone of APS-G2 is essential for superior bioactivity, contrasting with the diminished efficacy of the heterogeneous, branched APS-A1. APS-G2 significantly suppressed pro-inflammatory cytokines, elevated anti-inflammatory mediators, and improved intestinal barrier integrity by preventing DSS-induced apoptosis of intestinal epithelial cells and promoting the expression of tight junction proteins. The underlying mechanisms involve coordinated regulation of the SIRT1/PGC-1α/NF-κB pathway and FXR-mediated signaling. These findings elucidate the structure-function relationships of APS derivatives and suggest that enzymatic engineering can enhance the therapeutic potential of polysaccharides for IBD management.

Glycosylation, an effective strategy for enhancing the bioavailability and stability of dietary flavonoids, has primarily focused on developing enzyme resources from bacteria and plants. However, exploration of UDP glycosyltransferases (UGTs) from fungi remains insufficient. An in-depth study of fungal glycosyltransferases will enhance our understanding of the potential benefits of glycosylation in fungi and expand the range of available biocatalysts for glycosylation. In this study, we identified three key enzymes from Trametes hirsuta: UDP-glucose dehydrogenase (ThUGDH), UDP-xylose synthase (ThUXS), and UDP-flavonoid glycosyltransferase (Th3GT). These enzymes are involved in the synthesis of UDP sugar donors and the glycosylation of quercetin, efficiently converting it into UDP-GlcA, UDP-Xyl, quercetin-3-O-β-d-glucoside (Q3G), and quercetin-3-O-β-d-xylopyranoside (Q3X). Through mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy, we confirmed the structures of these compounds. We employed a simplified purification process to obtain high-purity sugar donors and quercetin glycosides. Notably, glycosylation reduces the cytotoxicity of quercetin while improving its capacity to scavenge reactive oxygen species (ROS). In summary, this study not only enriches the knowledge framework of fungal UGTs but also provides valuable enzyme resources for industrial enzyme-catalyzed synthesis of UDP sugar donors and flavonoid glycosides.

The cytosolic delivery of therapeutic proteins represents a promising strategy for addressing diseases caused by protein dysfunction. Despite significant advances, efficient delivery remains challenging due to barriers such as cell membrane impermeability, endosomal sequestration and protein instability. This review summarises recent progress in protein delivery systems, including physical, chemical and biological approaches, with a particular focus on strategies that enhance endosomal escape and targeting specificity. We further discuss the clinical translatability of these approaches and propose future directions for improving delivery efficiency and safety, ultimately unlocking the therapeutic potential of intracellular proteins.

Immunogenic cell death (ICD) is a promising approach for generating antitumor immune responses to treat patients with cancer. However, the stereotactic induction of ICD and spatiotemporal synchronized activation of the tumor-specific immune response pose two critical challenges. Here, we report the basic principles and systematic development of a modular metalloprotein platform designated the protease-activated PSTAGylated in situ tumor vaccine (PPTV). Ferritin was used as a vaccine framework, with its outer surface fused with an optimal mito-disrupt peptide and its inner cavity loaded with manganese ions (Mn2+). Moreover, a protease-activated PSTAGylated prodrug strategy was developed to circumvent the major issues associated with ferritin, such as liver interception and drug leakage. We also demonstrated the activation of prodrugs in tumor lysates from patients. In subcutaneous and orthotopic tumor transplantation mouse models, the PPTV effectively codelivered mito-disrupt peptides and Mn2+ into tumors, leading to a combined effect of ICD and cGAS-STING cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway activation, thereby achieving a potent antitumor immune response. The combined application of PPTV and anti-programmed death-ligand 1 resulted in the eradication of established tumors. These results underscore the applicability of the PPTV as an innovative in situ vaccine nanoplatform for establishing an antitumor immune response. This modular metalloprotein vaccine has clinical translational potential because of its simple generation and safety profile.

Tumor neoantigens, defined as tumor-specific antigens arising from somatic mutations, have shown great potential as targets for cancer vaccines in clinical studies. However, the number of neoantigens capable of effectively activating immune responses is quite limited. Over the past few decades, tumor neoantigen vaccines based on MHC-I epitopes that activate CD8+ T cells have been extensively studied. However, growing evidence suggests that CD4+ T cells are important in cancer immunotherapy. In contrast to CD8+ T cells, the receptors on CD4+ T cells exhibit a wider range of antigen peptide-MHC recognition, which can detect more tumor mutation antigens. In our earlier studies, a nitrated CD4+ T-cell epitope (NitraTh) was constructed as a novel CD4+ T-cell epitope that can enhance the immunogenicity of multiple tumor antigens. Therefore, we designed vaccines targeting MHC-II neoantigen epitopes using the nitrated T-cell epitope containing immunogenic amino acids. We found that vaccines conjugated with NitraTh exhibited enhanced immunogenicity. Crucially, the NitraTh-modified MHC-II tumor neoantigen vaccines increased the proportion of CD4+ T cells that infiltrate tumors and the spleen, elevated the expression of several cytokines with antitumor effects and facilitated the transformation of CD4+ T cells into Th1 cells, thereby reducing tumor growth. Additionally, the nitrated epitope has been shown to transform naïve CD4+ T cells into effector memory cells, thus facilitating enduring antitumor actions. The strategy of combining nitrated epitopes with MHC-II neoantigen epitopes confirms the significance of CD4+ T-cell immunity in cancer and may provide a novel approach for cancer vaccine design. SIGNIFICANCE STATEMENT: This study presents a novel design paradigm for tumor vaccines-combining MHC-II epitopes with nitrated CD4+ T-cell epitopes. This approach promotes the differentiation of CD4+ T cells toward a Th1 phenotype and generates long-lasting effector memory CD4+ T cells. Under the enhanced effects of CD4+ T cells, the vaccines we designed achieved superior antitumor efficacy and improved the immunosuppressive tumor microenvironment.

Diabetic encephalopathy (DE), characterized by cognitive impairment, currently lacks targeted treatment. Previous studies have shown that Sarcandra glabra extracted residue polysaccharide (SERP) exhibited hypoglycemic effects either in vitro or in streptozotocin-induced diabetes mice. However, the therapeutic effect of SERP on DE was not elucidated. This study investigated the therapeutic effect of SERP on DE and its underlying mechanism. Our results revealed that SERP regulates glucose and lipid metabolism, improves cognitive function, and exhibits diminished activity post-antibiotic intervention. Importantly, we discovered a novel mechanism by which SERP modulates the gut microbiota, specifically enriching Bacteroidales S24-7, resulting in elevated levels of butyric acid in the intestine. This regulation modulates the intestinal endocrine cell lipid metabolism level, restores damaged intestinal barriers and neural epithelial circuits, thus exhibiting cure effects. Our findings suggest that SERP could become a candidate for treating DE, potentially involving the regulation mechanism of the “microbiota-gut-brain axis”. This study underscores the unique therapeutic efficacy of SERP in managing DE, offering fresh drug candidates and innovative treatment strategies for this challenging condition.

Background and PurposeBecause of the absence of effective therapies for metabolic dysfunction-associated steatohepatitis (MASH), there is a rising interest in fibroblast growth factor 21 (FGF21) analogues due to their potential anti-fibrotic activities in MASH treatment. PsTag-FGF21, a long-acting FGF21 analogue, has demonstrated promising therapeutic effects in several MASH mouse models. However, its efficacy and mechanism against MASH-related fibrosis remain less well defined, compared with the specific mechanisms through which FGF21 improves glucose and lipid metabolism.Experimental ApproachThe effectiveness of PsTag-FGF21 was evaluated in two MASH-fibrosis models. Co-culture systems involving macrophages and hepatic stellate cells (HSCs) were employed for further assessment. Hepatic macrophages were selectively depleted by administering liposome-encapsulated clodronate via tail vein injections. RNA sequencing and cytokine profiling were conducted to identify key factors involved in macrophage-HSC crosstalk.Key ResultsWe first demonstrated the significant attenuation of hepatic fibrosis by PsTag-FGF21 in two MASH-fibrosis models. Furthermore, we highlighted the crucial role of macrophage phenotypic switch in PsTag-FGF21-induced HSC deactivation. FGF21 was demonstrated to regulate macrophages in a PsTag-FGF21-like manner. NR4A1, a nuclear factor which is notably down-regulated in human livers with MASH, was identified as a mediator responsible for PsTag-FGF21-induced phenotypic switch. Transcriptional control over insulin-like growth factor 1, a crucial factor in macrophage-HSC crosstalk, was exerted by the intrinsically disordered region domain of NR4A1.Conclusion and ImplicationsOur results have elucidated the previously unclear mechanisms through which PsTag-FGF21 treats MASH-related fibrosis and identified NR4A1 as a potential therapeutic target for fibrosis.

Altered glycosylation profiles have been correlated with potential drug targets in various diseases, including Alzheimer’s disease (AD). In this area, the linkage between bisecting N-acetylglucosamine (GlcNAc), a product of N-acetylglucosaminyltransferase III (GnT-III), and AD has been recognized, however, our understanding of the cause and the causative role of this aberrant glycosylation in AD are far from completion. Moreover, the effects and mechanisms of glycosylation-targeting interventions on memory and cognition, and novel targeting strategies are worth further study. Here, we showed the characteristic amyloid pathology-induced and age-related changes of GnT-III, and identified transcription factor 7-like 2 as the key transcription factor responsible for the abnormal expression of GnT-III in AD. Upregulation of GnT-III aggravated cognitive dysfunction and Alzheimer-like pathologies. In contrast, loss of GnT-III could improve cognition and alleviate pathologies. Furthermore, we found that an increase in bisecting GlcNAc modified ICAM-1 resulted in impairment of microglial responses, and genetic inactivation of GnT-III protected against AD mechanistically by blocking the aberrant glycosylation of ICAM-1 and subsequently modulating microglial responses, including microglial motility, phagocytosis ability, homeostatic/reactive state and neuroinflammation. Moreover, by target-based screening of GnT-III inhibitors from FDA-approved drug library, we identified two compounds, regorafenib and dihydroergocristine mesylate, showing pharmacological potential leading to modulation of aberrant glycosylation and microglial responses, and rescue of memory and cognition deficits.

Personalized neoantigen therapy has shown long-term and stable efficacy in specific patient populations. However, not all patients have sufficient levels of neoantigens for treatment. Although somatic mutations are commonly found in tumours, a significant portion of these mutations do not trigger an immune response. Patients with low mutation burdens continue to exhibit unresponsiveness to this treatment. We propose a design paradigm for neoantigen vaccines by utilizing the highly immunogenic unnatural amino acid p-nitrophenylalanine (pNO2Phe) for sequence alteration of somatic mutations that failed to generate neoepitopes. This enhances the immunogenicity of the mutations and transforms it into a suitable candidate for immunotherapy. The nitrated altered epitope vaccines designed according to this paradigm is capable of activating circulating CD8+ T cells and inducing immune cross-reactivity against autologous mutated epitopes in different MHC backgrounds (H-2Kb, H-2Kd, and human HLA-A02:01), leading to the elimination of tumour cells carrying the mutation. After immunization with the altered epitopes, tumour growth was significantly inhibited. It is noteworthy that nitrated epitopes induce tumour-infiltrating macrophages to differentiate into the M1 phenotype, surprisingly enhancing the MHC II molecule presenting pathway of macrophages. Nitrated epitope-treated macrophages have the potential to cross-activate CD4+ and CD8+ T cells, which may explain why pNO2Phe can enhance the immunogenicity of epitopes. Meanwhile, the immunosuppressive microenvironment of the tumour is altered due to the activation of macrophages. The nitrated neoantigen vaccine strategy enables the design of vaccines targeting non-immunogenic tumour mutations, expanding the pool of potential peptides for personalized and shared novel antigen therapy. This approach provides treatment opportunities for patients previously ineligible for new antigen vaccine therapy.