Enhancing RNA-lipid nanoparticle delivery: Organ- and cell-specificity and barcoding strategies

Journal of Controlled Release, Journal Year: 2024, Volume and Issue: 375, P. 366 - 388

Published: Sept. 18, 2024

Recent advancements in RNA therapeutics highlight the critical need for precision gene delivery systems that target specific organs and cells. Lipid nanoparticles (LNPs) have emerged as key vectors delivering mRNA siRNA, offering protection against enzymatic degradation, enabling targeted cellular uptake, facilitating cargo release into cytosol. This review discusses development optimization of organ- cell-specific LNPs, focusing on their design, mechanisms action, therapeutic applications. We explore innovations such DNA/RNA barcoding, which facilitates high-throughput screening precise adjustments formulations. address major challenges, including improving endosomal escape, minimizing off-target effects, enhancing efficiencies. Notable clinical trials recent FDA approvals illustrate practical applications future potential LNP-based therapies. Our findings suggest while considerable progress has been made, continued research is essential to resolve existing limitations bridge gap between pre-clinical evaluation safety efficacy therapeutics. highlights dynamic LNP research. It outlines a roadmap RNA-based medicine.

Language: Английский

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CAR-macrophages: tailoring cancer immunotherapy

Frontiers in Immunology, Journal Year: 2025, Volume and Issue: 15

Published: Jan. 14, 2025

The ability of effector immune cells to target and eliminate tumor by focusing on tumor-associated antigens is crucial for the success immunotherapy. Chimeric Antigen Receptor (CAR)-modified have revolutionized cancer immunotherapy, primarily with CAR-T showing remarkable in hematological cancers. Numerous cell-based therapies, such as cell, TIL, CAR-NK T-cell receptor (TCR)-based are currently undergoing clinical pre-clinical evaluation across various types. However, these therapies possess limitations, including their inability penetrate stoma, change TME, exaggerated inflammatory responses. To overcome this, developing advanced flexible strategies precisely while preserving homeostasis patients imperative. One promising approach involves using engineered macrophages (TAMs), which plastic nature constitute approximately 50% microenvironment (TME) indispensable both progression regression. CAR-macrophages aim reprogram toward M1 TAM phenotype enable them immune-suppressive TME facilitate immune-mediated destruction tumors. Body textThe typically comprises cells, more than total cell mass play a role or Tumor-associated extremely can adapt function response environmental stimuli. TAMs within exist two distinct phenotypes: (regulatory) M2 (trophic). produces proinflammatory cytokines like IL-12 TNF-α, activates T attack demonstrate antitumor activity. presence tumors often linked better outcomes due regulate growth. advances, intrinsic extrinsic factors drive polarization tumor-infiltrating into [1], supports development. These M2-polarized secrete pro-tumoral angiogenic factors, IL-10, TGF-β, VEGF, contributing an immunosuppressive environment that favors growth [2]. Various efforts under progress engineering express receptors recognize vide range (TAA) pipeline, allowing selectively destroy cells. macrophages, known Macrophages (CAR-Macrophages) capable enough priming reactive T-cells keeping natural tumor-targeting [3]. Thus, it clear CAR-Macrophage (CAR-M) therapy would be potential asset immunotherapy many solid Underlying MechanismCAR-Macrophage recognizes specific through its chimeric antigen (CAR), leading enhanced phagocytosis concomitant presentation subsequent activation. CAR-Ms modulate secreting cytokines, metalloproteinases (MMPs), Reactive oxygen species (ROS) serine proteases altogether infiltration anti-tumor responses Each mechanisms (Figure 1) contributes overall effectiveness CAR-Macrophages targeting eliminating making therapeutic CAR-engineered were shown actively migrate sites, could locally deliver and/or cytotoxic substances antigen-specific environments when used drug delivery system, significantly alter [4]. leverage tumor-homing tendencies cargo induce activity niche. With help regenerative technologies, induced pluripotent stem (iPSCs) make CD19 / mesothelin+ M1programmed bear strong anti-tumoral [5]. Engineered derived from diverse sources, primary human monocytes/macrophages, (iPSCs), hematopoietic progenitor (HSPCs). Abdin et al. (2023) iPSCs, HSPCs generate anti-CD19 CAR-Ms. They generated αCD19 CAR constructs lentiviral vectors verified via sequencing. CD34+ isolated cord blood, transduced, differentiated macrophages. iPSCs cultured, mesoderm-primed, defined cytokines. Cancer lines patient-derived samples transduced co-cultured assess phagocytosis. Then they flow cytometry, confocal microscopy, western blotting verify action displayed CD19+ pro-inflammatory responses, adaptive recruitment, scRNA sequencing revealed activation pathways cytokine upregulation [6]. In another study Zhang (2023), utilized CRISPR-Cas9 gene editing integrate anti-GD2 AAVS1 locus (hPSCs). developed serum- feeder-free differentiation protocol (CAR-Ms) arterial endothelial-to-hematopoietic transition (EHT). this way demonstrated potent against GD2-expressing neuroblastoma melanoma vitro vivo. It useful generating off-the-shelf CAR-Ms, advancing applications [7]. improve efficiency functionality hPSC shen (2024) optimized monolayer-based system achieving stable expression tumoricidal vitro. address diminished vivo activity, employed interferon-γ monophosphoryl lipid-A innate activation, repolarizing hPSC-CAR-Ms Additionally, activating collaborative innate-adaptive amplifying effects [8]. Using high-throughput screening, Mukalel identified oxidized lipid nanoparticles (oLNPs) monocyte tropism effective mRNA delivery. C14-O2 oLNP successfully CD19-CAR monocytes vivo, significant B depletion, highlighting [9]. Similarly, numerous novel approaches being developed, efficacy existing methods continues undergo constant refinement enhancement. ongoing advancements precision, scalability, current ensuring meet evolving needs applications, particularly fields cellular. Reprogramming macrophage-directed strategy delivering [10] innovative improving rejection [11]. past, we amply returned/conditioned syngeneic donor mice dictated promoted high-grade highly invasive neuroendocrine pancreas. Most intriguingly, adoptive transfer reprogrammed (which regarded surrogate CAR-M) potentially normalized vasculature aided PanNETs [12-14]. On basis believe HER2 CD47-specific M, sensing clearing dead containing neighboring (immune) also skewed Th1 lymphocytes (CD8+ cells), secretion major effectors viz IFN-γ & IL-2, neutralizing exhaustion markers PD-1, TIGIT, LAG-3 [15]. This explained entitle niche [16]. mechanism might responsible HER2-directed CAR-macrophage (CT-0508) so far phase-1 trial (NCT04660929). interestingly was safe tolerable variety indicated CAR-M based raised hope large future. Challenges ProspectiveAlthough CAR-M-based several advantages however few bottlenecks still associated them. issues do not proliferate post-administration, amount tolerate limited, reduce interventions. Moreover, exogenous tend accumulate liver after passing lungs, neutralize potentials. complexity over murine model factor possesses challenge effective. Such heterogeneous restrict wide antigens, impact recognition/binding TSA/TAA –M TME. whether support local tumor-supportive should carefully addressed, especially at research stage. problem has been observed likely pose obstacle general. Both stability durability genetic modifications hurdle off-target stabilized CRISPR-based approach. prevent excessive inflammation, paramount maximizing therapies. achieved floxing iNOS Arginase CD47 promoter affording palliative iNOsFloxed/ArginaseFloxedCD47+CAR-M adverse impact. would, only enhance but ensure successful trials employ CAR-M. fidelity CAR–M program pave new patients.

Language: Английский

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Nanoparticle Targeting Strategies for Lipid and Polymer‐Based Gene Delivery to Immune Cells In Vivo

Small Science, Journal Year: 2024, Volume and Issue: unknown

Published: July 30, 2024

Lipid nanoparticles and polymeric are promising biomaterial platforms for robust intracellular DNA mRNA delivery, highlighted by the widespread use of nanoparticle‐ (NP) based vaccines to help end COVID‐19 pandemic. Recent research has sought adapt this nanotechnology transfect engineer immune cells in vivo. The system is an especially appealing target due its involvement many different diseases, ex vivo‐engineered cell therapies like chimeric antigen receptor (CAR) T therapy have already demonstrated remarkable clinical success certain blood cancers. Although gene delivery can potentially address some cost manufacturing concerns associated with current autologous therapies, transfecting vivo challenging. Not only extrahepatic NP lymphoid organs difficult, but particular resistance transfection. Despite these challenges, modular nature NPs allows researchers examine critical structure–function relationships between a particle's properties ability specifically Herein, several nanomaterial components outlined, including targeting ligands, nucleic acid cargo, chemical properties, physical route administration optimal

Language: Английский

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Better, Faster, Stronger: Accelerating mRNA‐Based Immunotherapies With Nanocarriers

Wiley Interdisciplinary Reviews Nanomedicine and Nanobiotechnology, Journal Year: 2024, Volume and Issue: 16(6)

Published: Nov. 1, 2024

ABSTRACT Messenger ribonucleic acid (mRNA) therapeutics are attracting attention as promising tools in cancer immunotherapy due to their ability leverage the vivo expression of all known protein sequences. Even small amounts mRNA can have a powerful effect on vaccines by promoting synthesis tumor‐specific antigens (TSA) or tumor‐associated (TAA) antigen‐presenting cells (APC). These then presented T cells, eliciting strong antitumor immune stimulation. The potential be further enhanced expressing immunomodulatory agents, such cytokines, antibodies, and chimeric antigen receptors (CAR), enhancing tumor immunity. Recent research also explores mRNA‐encoded death inducers microenvironment (TME) modulators. Despite its promise, clinical translation mRNA‐based anticancer strategies faces challenges, including inefficient targeted delivery vivo, failure endosomal escape, inadequate intracellular release, resulting poor transfection efficiencies. Inspired approval lipid nanoparticle‐loaded against coronavirus disease 2019 (COVID‐19) encouraging outcomes therapies trials, innovative nonviral nanotechnology systems been engineered. aim advance immunotherapies from application. This review summarizes recent preclinical progress polymeric nanomedicines for delivering therapeutics, cytokines antibody‐based immunotherapies, vaccines, CAR therapies. It addresses advanced direct oncolysis TME reprogramming highlights key challenges translating these use, exploring future perspectives, role artificial intelligence machine learning development.

Language: Английский

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Metabolic Reprogramming of Cancer‐Associated Fibroblasts: Transforming Tumor Accomplices into Immunotherapeutic Allies

Advanced Functional Materials, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 9, 2024

Abstract Cancer‐associated fibroblasts (CAFs) play a pivotal role in the metabolic symbiosis that drives tumor proliferation and immune evasion. Paradoxically, eliminating CAFs can disrupt tissue homeostasis potentially accelerate spread. To address this challenge, study proposes novel strategy to reprogram CAFs, transforming them into “anchors” “fuel stations” for anti‐tumor cells, thereby enhancing response against tumors. Utilizing fibroblast‐specific lipid nanoparticle (LNP) delivery system, key genes are targeted downregulated, specifically hexokinase 2 (HK2) mitochondrial cytochrome c oxidase I (MTCO1). The dual inhibition of glycolysis respiration consequently results glucose overload dysfunction. As result, these energy‐deprived exhibit high expression MHC II molecules inflammatory cytokines, promoting cell infiltration providing essential fuel subsequent activation proliferation. Furthermore, reprogramming reduced angiogenesis an microenvironment characterized by M1 macrophage polarization enhanced lymphocyte infiltration. Consequently, approach improves efficacy checkpoint inhibitors (ICIs) castration‐resistant prostate cancer (CRPC). Thus, allies offers promising overcome limitations current ICIs therapies.

Language: Английский

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Pooled nanoparticle screening using a chemical barcoding approach

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 26, 2024

We report the development of a small molecule-based barcoding platform for pooled screening nanoparticle delivery. Using aryl halide-based tags (halocodes), we achieve high-sensitivity detection via gas chromatography coupled with mass spectrometry or electron capture. This enables and tracking nanoparticles minimal halocode concentrations without altering their physicochemical properties. To demonstrate utility our screening, synthesized halocoded library polylactide-co-glycolide (PLGA) quantified uptake in ovarian cancer cells manner. Our findings correlate conventional fluorescence-based assays. Additionally, potential halocodes spatial mapping using imaging (MSI). Halocoding presents an accessible modular capable quantifying delivery nanocarrier libraries range biological settings.

Language: Английский

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Pharmaceutical Strategies for Optimized mRNA Expression

Biomaterials, Journal Year: 2024, Volume and Issue: 314, P. 122853 - 122853

Published: Sept. 27, 2024

Language: Английский

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