Citations

Cancer immunotherapy is a transformative technology allowing for the elimination of malignant cells through immune system stimulation; however, the large-scale clinical application of adoptive cellular therapy technologies is limited due to difficulties in ex vivo immune cell engineering, high costs, and complicated manufacturing process. Recently, in vivo immune cell engineering was discovered as an emerging approach that allows for the genetic reprogramming of immune cells in patients, thus circumventing many problems inherent to traditional cellular therapy technologies. Breakthroughs in targeted drug delivery systems such as cell-penetrating peptides, antibody- and receptor-mediated delivery approaches, lipid nanoparticles (LNPs), polymeric drug carriers, and biomimetic delivery systems allow for the more effective selective delivery of messenger RNA (mRNA), clustered regularly interspaced short palindromic repeats (CRISPR)/Cas gene editing systems, proteins, and other types of cargo to the target immune cells. In vivo immune cell engineering allows for the efficient generation of chimeric antigen receptor T-cells (CAR-T), CAR- natural killer (NK) cells, engineered macrophages, dendritic cells, and other immune cell types; moreover, transient and permanent immune reprogramming is achievable via vivo technologies. The present review discusses the biological barriers limiting immune cell-targeted delivery, including biodistribution, cellular uptake, intracellular transport, and endosomal escape of delivered drugs, and reviews the latest advancements in targeted delivery technologies addressing these obstacles. We also address the latest applications of in vivo immune cell engineering in hematological malignancies and solid tumors, including mRNA therapeutics, CRISPR-based genome editing, and advanced programmable immunotherapy technologies. Furthermore, we discuss unresolved issues related to the specificity of targeting, efficiency of drug delivery, safety of treatment, manufacturing processes, and regulation of in vivo immune engineering technologies and outline future opportunities arising from the integration of nanomedicine, synthetic biology, genome engineering, and artificial intelligence-assisted materials science.

Introduction Recent transcriptome analysis has demonstrated increased expression of Vascular Endothelial Growth Factor receptor-1 (VEGFR-1/FLT1) and in AD brain. Increased expression of VEGFR1 and its ligand VEGFB were associated with a more rapid rate of cognitive decline, providing evidence of a potential link between increased VEGFR-1 expression in AD pathogenesis. In this study, we explored the potential role of VEGFR-1 expression in neurons on AD pathology. Methods To confirm VEGFR1 expression in AD brains, we first performed immunostaining in AD brain sections (AD - Braak stage V-VI, and normal controls - Braak 0-II). And to determine a potential detrimental role of neuronal VEGFR1 expression on AD associated pathologies, we exposed SH-SY5Y human neuroblastoma cells and mouse primary neurons to either hypoxia conditions (1%O 2 ) or 5 μ Aβ1-42 oligomers or fibrils for 24, 28 and 72hrs. Results In this study, we found preferential staining of VEGFR-1 in the neuropil and neuronal cell bodies both in AD and Control hippocampus and increased VEGFR-1 immunoreactivity in dystrophic neuritic processes in the vicinity of Thio-S positive amyloid plaques in AD brains. And treatment of SH-SY5Y human neuroblastoma cell line and mouse primary neurons, with either hypoxia conditions or Aβ1-42 oligomers, resulted in increased VEGFR-1 expression and cleaved caspase 3 activation, leading to neuronal toxicities/cell death. Similarly, treatment with Aβ1-42 fibrils also increased VEGFR-1 and cleaved caspase 3 protein levels in the SH-SY5Y cells whereas treatment with Aβ1-42 monomers had no effect on VEGFR-1 expression. In addition, we show that over-expression of VEGFR-1 intracellular domains in SH-SY5Y cells directly induced neuronal toxicities and importantly, siRNA-mediated knockdown of VEGFR-1 in neurons prevented the hypoxia, Aβ1-42 oligomer and Aβ1-42 fibril-induced toxicities and cell death phenotypes. Treatment with either hypoxia or Aβ1-42 oligomers also reduced expression of cell survival genes including VEGFR-2 and Hippo pathway YAP1 and siRNA-mediated VEGFR-1 knockdown in the neurons normalized expression of both VEGFR-2 and YAP1. Using differential gene expression analysis, we demonstrated upregulation of several inflammatory/interferon-stimulated genes (ISGs) as well as increased expression of genes involved in activation of oxidative stress and cell death pathways in response to Aβ1-42 oligomers treatment in mouse primary neurons. And siRNA-mediated VEGFR-1 knockdown in the mouse primary neurons, reduced gene expression of both the ISGs and oxidative stress/cell death pathways in response to Aβ1-42 oligomer treatment. Discussion In summary, these results show that siRNA-mediated knockdown of VEGFR-1 in neurons significantly prevented hypoxia, Aβ1-42 oligomer and Aβ1-42 fibril-induced cellular toxicities and cell death phenotypes, indicating a potential detrimental role of aberrant VEGFR-1 expression and signaling in response to AD associated pathologies.

Tumor Treating Fields (TTFields) represent a novel, non-invasive therapeutic modality in oncology that employs low-intensity, intermediate-frequency alternating electric fields to disrupt mitotic processes and induce cancer cell death. This review integrates mechanistic, preclinical, and emerging clinical evidence supporting the integration of TTFields with immunotherapeutic strategies in pancreatic ductal adenocarcinoma (PDAC). Although immunotherapy has transformed the treatment landscape across multiple malignancies, its efficacy in PDAC remains limited due to the tumor’s dense stroma, immunosuppressive microenvironment, and low immunogenicity. Preclinical investigations suggest that TTFields may potentiate immune-based therapies by enhancing antigen presentation, modulating the tumor microenvironment (TME), and attenuating mechanisms of immune resistance. We highlight studies evaluating TTFields in combination with immune checkpoint inhibitors (ICI), adoptive cellular therapies, and cancer vaccines, emphasizing their potential synergistic effects in PDAC. Clinically, the phase II PANOVA-2 trial demonstrated feasibility and encouraging survival outcomes with TTFields in combination with gemcitabine and nab-paclitaxel, providing the rationale for the ongoing phase III PANOVA-3 trial and the phase II PANOVA-4 trial, which combines TTFields with chemotherapy and atezolizumab. Additional clinical experiences in glioblastoma (GBM) and non-small-cell lung cancer (NSCLC) further substantiate the broader applicability of TTFields as an immunomodulatory adjunct. Remaining challenges include optimizing treatment sequencing, identifying predictive biomarkers, and managing TTFields-associated toxicities. Collectively, current evidence positions TTFields as a promising strategy to augment immunotherapy in PDAC, warranting further translational and clinical investigation to establish its role in reshaping therapeutic paradigms.

With limited therapeutic progress despite aggressive multimodal treatment, glioblastoma (GBM) remains one of the deadliest primary brain tumors. Emerging evidence suggests that GSCs are key drivers of tumor initiation, intratumoral heterogeneity, therapeutic resistance, and recurrence. GSCs retain self-renewal capacity, multilineage differentiation potential, and remarkable plasticity, enabling them to adapt to diverse microenvironmental cues. These properties are upheld by dysregulated developmental and oncogenic signaling pathways, including Notch, Wnt/β-catenin, Hedgehog, PI3K/AKT/mTOR, and STAT3, as well as epigenetic and metabolic reprogramming. In addition, dedicated niches such as hypoxic and perivascular microenvironments critically support GSC maintenance and immune evasion. In this review, we summarize the current understanding of the molecular pathways governing GSC biology, examine their interactions with the tumor microenvironment, and discuss emerging therapeutic strategies targeting GSCs, including pathway inhibition, differentiation therapy, immunotherapy, and nanomedicine-based drug delivery. We highlight key challenges and future directions for translating GSC-targeted approaches into effective clinical interventions for GBM.

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) is driven predominantly by oncogenic KRAS signaling and characterized by an immunosuppressive, fibrotic tumor microenvironment that limits therapeutic responses. RMC-6236, a pan-KRAS inhibitor, and ONIOYID, an investigational anti-tumor agent, each demonstrate partial activity in KRAS-mutant cancers. However, their combined therapeutic potential in PDAC remains unexplored. Here, we evaluated the anti-tumor efficacy, survival benefit, and immunomodulatory effects of RMC-6236, ONIOYID and irinotecan alone and dual combinations multiple in vitro and in vivo PDAC models. Methods: In vitro cytotoxicity assays were performed using KPC, PANC-1, AsPC-1, and PANC-02 cell lines. In vivo therapeutic studies were conducted in orthotopic KPC and PANC-1 tumor models. RMC-6236 was administered orally at 10 mg/kg; ONIOYID and irinotecan was administered intravenously at 5 mg/kg. Tumor growth inhibition, survival, collagen deposition, and immune cell infiltration were analyzed. Tumor microenvironment remodeling was quantified via immunohistochemistry for CD8, CD4, collagen I, and fibronectin. Results: The combination of RMC-6236 and ONIOYID elicited potent anti-tumor effects across all evaluated PDAC cell lines, achieving greater than 90% growth inhibition in vitro. In vivo, co-treatment resulted in more than 90% tumor regression in both KPC and PANC-1 xenograft models, markedly surpassing the efficacy of individual monotherapies and other dual-agent controls. Combination therapy significantly extended overall survival and produced sustained suppression of tumor progression. Mechanistic analyses revealed a substantial reduction in collagen and fibronectin deposition within the tumor microenvironment, consistent with attenuation of desmoplasia. Immune profiling demonstrated a robust enhancement of intratumoral CD8+ and CD4+ T-cell infiltration, along with a pronounced decrease in survivin expression. Importantly, no observable systemic toxicity was detected in treated animals. Conclusions: RMC-6236 combined with ONIOYID yields synergistic anti-tumor effects, remodels the fibrotic PDAC microenvironment, enhances T-cell infiltration, and significantly improves survival in aggressive pancreatic tumor models. These findings provide strong rationale for clinical evaluation of RMC-6236-based combination therapies for KRAS driven PDAC. Citation Format: Hari Krishnareddy Rachamala, Fang Wei, Debabrata Mukhopadhyay, Hani M. Babiker. Combined KRAS pathway inhibition and ONIOYID treatment enhances tumor regression, attenuates desmoplasia, and augments T cell infiltration in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5842.

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) remains highly lethal due to aggressive biology, stromal desmoplasia, and profound immunotherapy resistance. Tumor Treating Fields (TTFields) disrupt mitosis and induce immunogenic cell death (ICD), type I interferon signaling, and microenvironment remodeling, providing a rationale for combining TTFields with immune checkpoint inhibition. This is the first systematic review to comprehensively synthesize mechanistic and clinical evidence for TTFields immunotherapy combinations across solid tumors, with emphasis on their translation relevance to PDAC. Methods: A systematic search of PubMed, Scopus, Web of Science, Embase, Cochrane Library, and ClinicalTrials.gov was conducted through October 26, 2025, following PRISMA 2020 guidelines. Eligible studies evaluated TTFields combined with immunotherapy. Two reviewers independently screened and extracted data using Covidence. Evidence was summarized narratively by tumor type, study design, and immunotherapeutic approach. Results: Thirty-seven studies met criteria, including preclinical and clinical investigations across glioblastoma, NSCLC, mesothelioma, PDAC, and other tumors. Preclinical models consistently showed that TTFields trigger ICD hallmarks such as calreticulin exposure, HMGB1 and ATP release, and activate STING/type I interferon pathways, enhancing dendritic-cell maturation and T-cell and NK-cell function. Across in vivo models, TTFields combined with PD-1 or CTLA-4 blockade, vaccines, or other immunotherapies reduced tumor growth and increased lymphocyte infiltration. Clinical evidence, although early-phase and heterogeneous, indicates the combination is feasible, with toxicity dominated by low-grade skin reactions and no excess immune-related adverse events. In glioblastoma, adding TTFields to pembrolizumab and temozolomide showed numerically improved survival. The LUNAR phase 3 trial in metastatic NSCLC demonstrated an overall survival advantage when TTFields were added to systemic therapy, including immunotherapy. Evidence in PDAC remains preliminary, but the ongoing PANOVA-4 study combining TTFields with atezolizumab and chemotherapy highlights emerging translational relevance. Conclusions: TTFields enhance antitumor immunity when combined with immunotherapy across solid tumors. In PDAC, these mechanisms may help counteract the immunosuppressive microenvironment. Ongoing gastrointestinal trials will clarify optimal combinations, biomarkers of response, and the clinical role of TTFields-based immunomodulation. Citation Format: Douaa Albelal, Hari Krishnareddy Rachamala, Hani Babiker. Synergistic immunomodulation by Tumor Treating Fields combined with immunotherapy across solid tumors: A systematic review with translational relevance to pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 3813.

Lipid-based delivery systems (LDS), including lipid nanoparticles (LNPs) and liposomes, have become indispensable tools in modern biomedicine owing to their biocompatibility, capacity to encapsulate diverse therapeutic agents, and potential for targeted delivery. Despite their clinical success, conventional batch-based manufacturing methods are hindered by variability, limited scalability, and complex processing steps, slowing their broader translation. Microfluidic technologies offer a transformative solution by enabling precise fluid handling, rapid mixing, and reproducible production of LDS with tunable physicochemical attributes such as particle size, lamellarity, and drug-loading efficiency. This review highlights advances in microfluidic design strategies, including hydrodynamic flow focusing, staggered herringbone mixers, and toroidal micromixers, and evaluates how critical parameters such as flow rate, solvent composition, and lipid concentration influence LDS performance. Furthermore, we discuss the application of microfluidics in drug delivery, nucleic acid therapeutics, and vaccine platforms, underscoring its role in improving scalability, quality control, and clinical translation. Finally, we examine current challenges, including throughput limitations and solvent handling, while outlining future directions for integrating emerging materials and additive manufacturing to optimize LDS fabrication. Collectively, microfluidic platforms provide a promising pathway for next-generation lipid nanomedicines with enhanced precision, reproducibility, and therapeutic efficacy.

ABSTRACT The standard treatment for Haemophilia B (HB) involves frequent intravenous (IV) infusions of recombinant factor IX (FIX), which imposes a significant financial burden on patients. FIX delivery as messenger RNA using lipid nanoparticles (LNPs) presents a promising alternative, but achieving prolonged and effective protein expression remains challenging. To this end, we developed Galactosylated Lipid Nucleic Acid complex (GaLiNAx) to enhance the delivery of chemically modified FIX mRNAs. The nanocarrier encompasses an ionizable galactosylated lipid to enable receptor‐mediated endocytosis and a transfection‐competent cationic lipid for optimal mRNA complexation. We incorporated truncated albumin UTRs and base modifications in functional FIX mRNA to enhance hepatic translation. In an HB mouse model, GaLiNAx demonstrated therapeutically relevant FIX protein expression for about one month after single dose offering a cost‐effective option. Safety assessments indicated that multiple doses of GaLiNAx could be administered without toxicity to any major organs. These results highlight the potential of long‐lasting mRNA therapy as an effective protein replacement strategy for HB, paving the way for future mRNA therapeutics targeting other monogenic liver disorders.