Spatiotemporal dynamic regulation of the CX3CL1-CX3CR1 axis: A double-edged sword in the tumor immune microenvironment and new strategies for precision therapy.

Shaping immunotherapy through the tumor microenvironment: Translational perspectives.

Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide, underscoring the urgent need for novel therapeutic approaches beyond standard treatments. Nanovaccines are a revolutionary platform in cancer immunotherapy, providing improved antigen delivery, enhanced immune activation, and targeted tumor targeting. This review critically discusses the promise of nanovaccines in HCC therapy, specifically focusing on their ability to induce strong antitumor immune responses while avoiding systemic toxicity. Key nanoplatforms such as lipid-based nanoparticles, polymeric carriers, and dendrimers are explained in detail to describe their mechanisms for encapsulating adjuvants and tumor-associated antigens to enhance immunogenicity. Further, we explain underlying mechanisms of action such as antigen cross-presentation, T-cell activation, and regulating the tumor microenvironment for immune evasion in HCC. Possible future directions in preclinical and clinical research and issues involving large-scale manufacturing and deployment are discussed, including combinatorial approaches using immune checkpoint inhibitors. This paper emphasizes the potentially transformative role of nanovaccines in the therapeutic regimen of HCC. It offers critical feedback on how their efficacy, safety, and translatability can be maximized for designing the next generation of cancer immunotherapies.

Recent advancements at the intersection of nanotechnology, artificial intelligence (AI), and immunotherapy are transforming the field of cancer treatment. This review explores the transformative potential of AI-guided nanorobots for in vivo immune cell programming. This strategy overcomes the limitations of conventional ex vivo adoptive cell therapies, including high cost, limited scalability, and reduced efficacy in solid tumors. We examine the principles of immune cell engineering, including CAR-T, CAR-NK, and TCR therapies, as well as the associated clinical challenges. Furthermore, we discuss how AI-guided nanorobots can autonomously navigate biological systems to deliver genetic or immunomodulatory payloads, remodel the tumor microenvironment, and enhance therapeutic precision. By integrating multimodal sensing and real-time decision-making capabilities, these nanorobots represent a novel class of autonomous agents that can detect cancer early, activate the immune system, and potentially intervene preemptively. This convergence of disciplines signals a new frontier in personalized, minimally invasive cancer therapy, offering hope for broader accessibility and improved outcomes. This review outlines a transformative approach to autonomous, minimally invasive, and personalized immune modulation, providing a blueprint for the next generation of cancer immunotherapy.

Nanocarrier vaccines represent a groundbreaking advancement in cancer immunotherapy, leveraging nanotechnology to enhance vaccine efficacy and specificity. This review examines the latest advancements in nanocarrier-based cancer vaccine formulations, focusing on the types of nanocarriers utilized and the critical role of their physicochemical properties in influencing immune responses. Key nanocarriers include liposomes, polymeric nanoparticles, lipid nanoparticles, self-assembled protein nanoparticles, inorganic nanoparticles, and virus-like particles. These nanocarriers improve antigen stability, protect against degradation, enable controlled release, and enhance uptake by antigen- presenting cells, resulting in stronger and more durable immune responses. The physicochemical characteristics of nanocarriers, including dimensions, form, surface charge, hydrophobicity, and degradability significantly influence vaccine efficacy by affecting cellular uptake, lymphatic trafficking, antigen presentation, and immune activation. Recent advancements optimize nanocarrier formulations to enhance antigen retention, immune interactions, and tumour modulation. Integrating immune-stimulatory agents like toll-like receptor agonists and cytokines, boosts immunogenicity, overcoming immune tolerance and improving outcomes. The emergence of new patents in nanocarrier- based cancer vaccines highlights innovative approaches in antigen stabilization, adjuvant selection, and targeted delivery. These patented technologies are driving the next generation of cancer immunotherapies, offering promising strategies for achieving precise, effective, and personalized cancer treatment. By synthesizing the latest findings, this review acts as a crucial reference for researchers and clinicians committed to progressing cancer nano vaccine innovation and understanding the emergence of new patents.

Oncolytic viruses (OVs) have the ability to efficiently enter, replicate within, and destroy cancer cells. This capacity to selectively target cancer cells while inducing long-term anti-tumor immune responses, makes OVs a promising tool for next-generation cancer therapy. Immunogenic cell death (ICD) induced by OVs initiates the cancer-immunity cycle (CIC) and plays a critical role in activating and reshaping anti-cancer immunity. Genetic engineering, including arming OVs with cancer cell-specific binders and immunostimulatory molecules, further enhances immune responses at various stages of the CIC, improving the specificity and safety of virotherapy.The aim of this study is to update current knowledge in immunotherapy using OVs and to highlight the remarkable plasticity of viruses in shaping the tumor immune microenvironment, which may facilitate anti-cancer treatment through various approaches. Research articles, meta-analyses, and systematic reviews were retrieved from PubMed, using the search terms ('Oncolytics' OR 'Immunotherapy' OR 'Virotherapy' OR 'Viral vector') AND 'gene therapy', without language restrictions. In this review, we discuss current strategies aimed at increasing the tumor specificity of OVs and improving their safety. We summarize and functionally categorize different biochemical approaches, with a focus on virus engineering and advancements in immunotherapy. Transduction targeting methods (e.g., xenotype switching, pseudotyping, cell receptor targeting) and non-transduction modifications (e.g., miRNA, optogenetics, transcriptional targeting) are critically reviewed. We also examine the mechanisms of ICD and viral modifications that contribute to efficient cancer cell death and modulation of cancer-specific immunity. Finally, we provide an outlook on promising future oncolytics and approaches with potential therapeutic benefit for the next generation of cancer immunotherapy. Immunogenic cell death induced by oncolytic viruses is a key mediator of potent anti-cancer immunity. The genetic integration of immunostimulatory molecules as regulatory elements into OV genomes significantly enhances their therapeutic potential, safety, and stability. Additionally, therapeutic potency can be further increased by deleting viral genes that inhibit apoptosis, thereby enhancing ICD. However, the synergistic effects of these modifications may vary significantly depending on the cancer type.

Current treatments for breast cancer (BC), particularly triple-negative BC, are limited in efficacy due to drug resistance and high recurrence rates. CD24, which is highly expressed in BCs and engages with its receptor Siglec-10/G (Siglec-10 in humans and Siglec-G in mice) on immune cells, represents a promising immune checkpoint blockade (ICB) target. As the engagement is mediated by a short signal transducer (ST) peptide displayed on the BC cell surface, targeting the peptide using antibodies has shown to be effective for BC treatment. Herein, an antibody-free approach is reported to achieve blockade of the CD24-Siglec-10/G signaling through the synthesis of signal transducer peptide-anchored nanoparticles (STNPs). The STNPs can effectively engage with macrophages, promoting enhanced phagocytosis of BC cells, triggering a broad immune response, and ultimately inhibiting tumor growth. The therapeutic effects can be further improved through encapsulation of RRx-001, a small molecule inhibitor of the CD47-SIRPα signaling. Compared to the antibody approach, the synthetic nanoparticle approach offers greater efficacy with lower side effects and enables combination therapy through a simple formulation. Moreover, the approach is versatile and could be adapted for targeting other ICB signaling, advancing the next generation of cancer immunotherapy.

Chimeric antigen receptor (CAR) cell therapies have revolutionized the field of cancer treatment with the breakthrough successes of CAR-T cell therapy in hematological malignancies. However, CAR-T cell efficacy in solid tumors has been limited by challenges such as the immunosuppressive tumor microenvironment (TME) and safety concerns like cytokine release syndrome (CRS) and graft-versus-host disease (GvHD). This review focuses on the latest advancements in CAR-NK cell and CAR-macrophage therapies, which offer potential solutions to these limitations. CAR-NK cells exhibit natural cytotoxicity and MHC-independent tumor targeting, providing a safer profile with lower risks of GvHD and CRS compared to CAR-T cells. Innovations in CAR-NK cell therapy includes structural optimizations, gene editing with CRISPR-Cas9, and the development of multispecific CARs, all contributing to enhanced anti-tumor efficacy, especially in solid tumors. Meanwhile, CAR-macrophages are effective in remodeling the TME through phagocytosis and immune activation, addressing the challenges of solid tumor infiltration. This review compares the mechanisms of action, clinical applications, and safety profiles of CAR-NK cells and CAR-macrophages, highlighting the synergistic potential of these therapies. Although clinical research on both therapies is still in its early stages, the promising preclinical and early clinical trial results underscore their potential as alternative immunotherapies for refractory cancers. Future research will focus on overcoming challenges in production scalability, improving in vivo persistence, and personalizing CAR therapies through precision medicine approaches. These two therapies will likely play pivotal roles in the next generation of cancer immunotherapies.

Immunotherapy is currently a standard of care in the treatment of many malignancies. However, predictable side effects caused by systemic administration of highly immunostimulatory molecules have been a serious concern within this field. Intratumoural expression or silencing of immunogenic and immunoinhibitory molecules using nucleic acid-based approaches such as plasmid DNA (pDNA) and small interfering RNA (siRNA), respectively, could represent a next generation of cancer immunotherapy. Here, we employed lipid nanoparticles (LNPs) to deliver either non-specific pDNA and siRNA, or constructs targeting two prominent immunotherapeutic targets OX40L and indoleamine 2,3-dioxygenase-1 (IDO), to tumours in vivo.In the B16F10 mouse model, intratumoural delivery of LNP-formulated non-specific pDNA and siRNA led to strong local immune activation and tumour growth inhibition even at low doses due to the pDNA immunogenic nature. Replacement of these non-specific constructs by pOX40L and siIDO resulted in more prominent immune activation as evidenced by increased immune cell infiltration in tumours and tumour-draining lymph nodes. Consistently, pOX40L alone or in combination with siIDO could prolong overall survival, resulting in complete tumour regression and the formation of immunological memory in tumour rechallenge models. Our results suggest that intratumoural administration of LNP-formulated pDNA and siRNA offers a promising approach for cancer immunotherapy.

The discovery of both cytotoxic T lymphocyte-associated antigen 4 (CTLA4) and programmed cell death protein 1 (PD1) as negative regulators of antitumour immunity led to the development of numerous immunomodulatory antibodies as cancer treatments. Preclinical studies have demonstrated that the efficacy of immunoglobulin G (IgG)-based therapies depends not only on their ability to block or engage their targets but also on the antibody's constant region (Fc) and its interactions with Fcγ receptors (FcγRs). Fc-FcγR interactions are essential for the activity of tumour-targeting antibodies, such as rituximab, trastuzumab and cetuximab, where the killing of tumour cells occurs at least in part due to these mechanisms. However, our understanding of these interactions in the context of immunomodulatory antibodies designed to boost antitumour immunity remains less explored. In this Review, we discuss our current understanding of the contribution of FcγRs to the in vivo activity of immunomodulatory antibodies and the challenges of translating results from preclinical models into the clinic. In addition, we review the impact of genetic variability of human FcγRs on the activity of therapeutic antibodies and how antibody engineering is being utilized to develop the next generation of cancer immunotherapies.

The cellular stress and immunity cycle is a cornerstone of organismal homeostasis. Stress activates intracellular and intercellular communications within a tissue or organ to initiate adaptive responses aiming to resolve the origin of this stress. If such local measures are unable to ameliorate this stress, then intercellular communications expand toward immune activation with the aim of recruiting immune cells to effectively resolve the situation while executing tissue repair to ameliorate any damage and facilitate homeostasis. This cellular stress-immunity cycle is severely dysregulated in diseased contexts like cancer. On one hand, cancer cells dysregulate the normal cellular stress responses to reorient them toward upholding growth at all costs, even at the expense of organismal integrity and homeostasis. On the other hand, the tumors severely dysregulate or inhibit various components of organismal immunity, for example, by facilitating immunosuppressive tumor landscape, lowering antigenicity, and increasing T-cell dysfunction. In this review we aim to comprehensively discuss the basis behind tumoral dysregulation of cellular stress-immunity cycle. We also offer insights into current understanding of the regulators and deregulators of this cycle and how they can be targeted for conceptualizing successful cancer immunotherapy regimen.

Abstract Engineered cell therapies, especially chimeric antigen receptor (CAR) T cell therapies, have shown much promise in treating various cancers, most notably hematologic malignancies. However, the success of these therapies is highly variable between cancer types and across patient populations. A growing body of literature suggests that certain T cell functional properties are key drivers of response; thus, it is imperative to develop a more thorough understanding of the functional heterogeneity in T cell populations in order to inform the design of the next generation of cancer immunotherapies. Here, we used a new technology our team developed known as nanovials, which are microfluidically manufactured hydrogel microparticles, to enable the specific activation and capture of secreted products of T cells on a single-cell level. Leveraging this platform, we characterized the secretion of IFN-γ and TNF-α by individual CD8 +T cells harvested from OT-I mice. After initial expansion after isolation, T cells were loaded into functionalized nanovials and specifically activated with OVA peptide. The cells loaded in the nanovials were then separated based on cytokine secretion levels via fluorescence-activated cell sorting (FACS). Unlike traditional cytokine secretion assays which require fixation and permeabilization, cells remain viable throughout our process, allowing for sorted subpopulations to be dissociated from the microparticles, expanded, and further analyzed via cytotoxicity, co-culture, or phenotypic studies. Collectively, our efforts introduce a new paradigm for elucidating the functional properties of immune cells, providing critical insights that will guide the future design of engineered cell therapeutics. M.K. is a recipient of a National Science Foundation Graduate Research Fellowship Program award.

Despite significant advances in cancer treatment, the metastatic spread of malignant cells to distant organs remains a major cause of cancer-related deaths. Natural killer (NK) cells play a crucial role in controlling tumor metastasis; however, the dynamics of NK cell-mediated clearance of metastatic tumors are not entirely understood. Herein, we demonstrate the cooperative role of NK and T cells in the surveillance of melanoma metastasis. We found that NK cells effectively limited the pulmonary seeding of B16 melanoma cells, while T cells played a primary role in restricting metastatic foci growth in the lungs. Although the metastatic foci in the lungs at the endpoint were largely devoid of NK cells, they played a prominent role in promoting T cell recruitment into the metastatic foci. Our data suggested that the most productive interaction between NK cells and metastatic cancer cells occurred when cancer cells were in circulation. Modifying the route of administration so that intravenously injected melanoma cells bypass the first liver passage resulted in significantly more melanoma metastasis to the lung. This finding indicated the liver as a prominent site where NK cells cleared melanoma cells to regulate their seeding in the lungs. Consistent with this notion, the liver and the lungs of the tumor-bearing mice showed dominance of NK and T cell activation, respectively. Thus, NK cells and T cells control pulmonary metastasis of melanoma cells by distinct mechanisms where NK cells play a critical function in shaping T cell-mediated in situ control of lung-seeded cancer cells. A precise understanding of the cooperative role of NK and T cells in controlling tumor metastasis will enable the development of the next generation of cancer immunotherapies.

Exosome transportation-mediated immunosuppression relief through cascade amplification for enhanced apoptotic body vaccination

Discovery and development of a novel N-(3-bromophenyl)-{[(phenylcarbamoyl)amino]methyl}-N-hydroxythiophene-2-carboximidamide indoleamine 2,3-dioxygenase inhibitor using knowledge-based drug design

In the past decade, immunotherapies have been emerging as an effective way to treat cancer. Among several categories of immunotherapies, immune checkpoint inhibitors (ICIs) are the most well-known and widely used options for cancer treatment. Although several studies continue, this treatment option has yet to be developed into a precise application in the clinical setting. Recently, omics as a high-throughput technique for understanding the genome, transcriptome, proteome, and metabolome has revolutionized medical research and led to integrative interpretation to advance our understanding of biological systems. Advanced omics techniques, such as multi-omics, single-cell omics, and typical omics approaches, have been adopted to investigate various cancer immunotherapies. In this review, we highlight metabolomic studies regarding the development of ICIs involved in the discovery of targets or mechanisms of action and assessment of clinical outcomes, including drug response and resistance and propose biomarkers. Furthermore, we also discuss the genomics, proteomics, and advanced omics studies providing insights and comprehensive or novel approaches for ICI development. The overview of ICI studies suggests potential strategies for the development of other cancer immunotherapies using omics techniques in future studies.

The discovery of immune checkpoint blockade has revolutionized the field of immuno-oncology and established the foundation for developing various new therapies that can surpass conventional cancer treatments. Most recent immunotherapeutic strategies have focused on adaptive immune responses by targeting T cell-activating pathways, genetic engineering of T cells with chimeric antigen receptors, or bispecific antibodies. Despite the unprecedented clinical success, these T cell-based treatments have only benefited a small proportion of patients. Thus, the need for the next generation of cancer immunotherapy is driven by identifying novel therapeutic molecules or new immunoengineered cells. To maximize the therapeutic potency via innate immunogenicity, the convergence of innate immunity-based therapy and biomaterials is required to yield an efficient index in clinical trials. This review highlights how biomaterials can efficiently reprogram and recruit innate immune cells in tumors and ultimately initiate activation of T cell immunity against advanced cancers. Moreover, the design and specific biomaterials that improve innate immune cells' targeting ability to selectively activate immunogenicity with minimal adverse effects are discussed.

Simple SummaryCD137 is an interesting immuno-oncology target. The recent advances in CD137 targeting technologies to mitigate toxicity while maintaining potency have made this space even more attractive. In this article, our current understanding of CD137 biology is reviewed along with data from clinical trials targeting T cell co-stimulatory receptors in the TNFRSF. Next generation CD137 targeting molecules currently in early clinical development are also reviewed. Finally, the future challenges of CD137 targeting molecules are discussed.Immune checkpoint inhibitors have altered the treatment landscape significantly in several cancers, yet not enough for many cancer patients. T cell costimulatory receptors have been pursued as targets for the next generation of cancer immunotherapies, however, sufficient clinical efficacy has not yet been achieved. CD137 (TNFRSF9, 4-1BB) provides co-stimulatory signals and activates cytotoxic effects of CD8+ T cells and helps to form memory T cells. In addition, CD137 signalling can activate NK cells and dendritic cells which further supports cytotoxic T cell activation. An agonistic monoclonal antibody to CD137, urelumab, provided promising clinical efficacy signals but the responses were achieved above the maximum tolerated dose. Utomilumab is another CD137 monoclonal antibody to CD137 but is not as potent as urelumab. Recent advances in antibody engineering technologies have enabled mitigation of the hepato-toxicity that hampered clinical application of urelumab and have enabled to maintain similar potency to urelumab. Next generation CD137 targeting molecules currently in clinical trials support T cell and NK cell expansion in patient samples. CD137 targeting molecules in combination with checkpoint inhibitors or ADCC-enhancing monoclonal antibodies have been sought to improve both clinical safety and efficacy. Further investigation on patient samples will be required to provide insights to understand compensating pathways for future combination strategies involving CD137 targeting agents to optimize and maintain the T cell activation status in tumors.

Tumor‐associated macrophages (TAMs) promote the immune suppressive microenvironment inside tumors and are, therefore, considered as a promising target for the next generation of cancer immunotherapies. To repolarize their phenotype into a tumoricidal state, the Toll‐like receptor 7/8 agonist imidazoquinoline IMDQ is site‐specifically and quantitatively coupled to single chain antibody fragments, so‐called nanobodies, targeting the macrophage mannose receptor (MMR) on TAMs. Intravenous injection of these conjugates result in a tumor‐ and cell‐specific delivery of IMDQ into MMRhigh TAMs, causing a significant decline in tumor growth. This is accompanied by a repolarization of TAMs towards a pro‐inflammatory phenotype and an increase in anti‐tumor T cell responses. Therefore, the therapeutic benefit of such nanobody‐drug conjugates may pave the road towards effective macrophage re‐educating cancer immunotherapies.

Natural killer (NK) cells are innate lymphocytes recognized for their important role against tumor cells. NK cells expressing chimeric antigen receptors (CARs) have enhanced effector function against various type of cancer and are attractive contenders for the next generation of cancer immunotherapies. However, a number of factors have hindered the application of NK cells for cellular therapy, including their poor in vitro growth kinetics and relatively low starting percentages within the mononuclear cell fraction of peripheral blood or cord blood (CB). To overcome these limitations, we genetically-engineered human leukocyte antigen (HLA)-A− and HLA-B− K562 cells to enforce the expression of CD48, 4-1BBL, and membrane-bound IL-21 (mbIL21), creating a universal antigen presenting cell (uAPC) capable of stimulating their cognate receptors on NK cells. We have shown that uAPC can drive the expansion of both non-transduced (NT) and CAR-transduced CB derived NK cells by >900-fold in 2 weeks of co-culture with excellent purity (>99.9%) and without indications of senescence/exhaustion. We confirmed that uAPC-expanded research- and clinical-grade NT and CAR-transduced NK cells have higher metabolic fitness and display enhanced effector function against tumor targets compared to the corresponding cell fractions cultured without uAPCs. This novel approach allowed the expansion of highly pure GMP-grade CAR NK cells at optimal cell numbers to be used for adoptive CAR NK cell-based cancer immunotherapy.