bims-drudre 2022-10-23 papers

bims-drudre

Biomed News

on Targeted drug delivery and programmed release mechanisms

Issue of 2022–10–23
seventeen papers selected by
Ceren Kimna, Technical University of Munich

Injectable Immunotherapeutic Hydrogel Containing RNA-Loaded Lipid Nanoparticles Reshapes Tumor Microenvironment for Pancreatic Cancer Therapy.
Aggregation-Induced-Emission Photosensitizer-Loaded Nano-superartificial Dendritic Cells with Directly Presenting Tumor Antigens and Reversed Immunosuppression for Photodynamically Boosted Immunotherapy.
Probiotic Spore-Based Oral Drug Delivery System for Enhancing Pancreatic Cancer Chemotherapy by Gut-Pancreas-Axis-Guided Delivery.
Enzyme-Catalyzed One-Step Synthesis of Ionizable Cationic Lipids for Lipid Nanoparticle-Based mRNA COVID-19 Vaccines.
Nociceptor neurons direct goblet cells via a CGRP-RAMP1 axis to drive mucus production and gut barrier protection.
3D microengineered vascularized tumor spheroids for drug delivery and efficacy testing.
Drug Delivery in Plants Using Silk Microneedles.
Photoacoustic image-guided biomimetic nanoparticles targeting rheumatoid arthritis.
Remodelling of tumour microenvironment by microwave ablation potentiates immunotherapy of AXL-specific CAR T cells against non-small cell lung cancer.
A combinational chemo-immune therapy using outer membrane vesicles for enhanced cancer therapy by RGD targeting.
Triggered Self-Sorting of Peptides to Form Higher-Order Assemblies in a Living System.
Photoswitchable Microgel for Dynamic Macrophage Modulation.
Lipidoid nanoparticles increase ATP uptake into hypoxic brain endothelial cells.
Lactose azocalixarene drug delivery system for the treatment of multidrug-resistant pseudomonas aeruginosa infected diabetic ulcer.
Machine Learning Guides Peptide Nucleic Acid Flow Synthesis and Sequence Design.
Airway epithelial cell-specific delivery of lipid nanoparticles loading siRNA for asthma treatment.
Targeted delivery of an anti-inflammatory corticosteroid to Ly6C/G-positive cells abates severity of influenza A symptoms.

Nano Lett. 2022 Oct 17.

Injectable Immunotherapeutic Hydrogel Containing RNA-Loaded Lipid Nanoparticles Reshapes Tumor Microenvironment for Pancreatic Cancer Therapy.

Chao Gao, Keman Cheng, Yao Li, Ruining Gong, Xiao Zhao, Guangjun Nie, He Ren.

  Pancreatic cancer immunotherapy is becoming a promising strategy for improving the survival rate of postsurgical patients. However, the low response rate to immunotherapy suggests a low number of antigen-specific T cells and a high number of immunosuppressive tumor-associated macrophages in the pancreatic tumor microenvironment. Herein, we developed an in situ injectable thermosensitive chitosan hydrogel loaded with lipid-immune regulatory factor 5 (IRF5) mRNA/C-C chemokine ligand 5 (CCL5) siRNA (LPR) nanoparticle complexes (LPR@CHG) that reprogram the antitumoral immune niche. The LPR@CHG hydrogel upregulates IRF5 and downregulates CCL5 secretion, which contribute to a significant increase in M1 phenotype macrophages. Tumor growth is controlled by effective M1 phenotype macrophage that initiate T cell-mediated immune responses. Overall, the LPR@CHG hydrogel is expected to be a meaningful immunotherapy platform that can reshape the immunosuppressive tumor microenvironment and improve the efficacy of current pancreatic immunotherapies while minimizing systemic toxicity.

Keywords:  RNA delivery; TAMs; chitosan hydrogel; immune therapy; liposome nanoparticles

DOI:  https://doi.org/10.1021/acs.nanolett.2c01994
Adv Mater. 2022 Oct 18. e2208555

Aggregation-Induced-Emission Photosensitizer-Loaded Nano-superartificial Dendritic Cells with Directly Presenting Tumor Antigens and Reversed Immunosuppression for Photodynamically Boosted Immunotherapy.

Zhihong Sun, Jie Liu, Yueying Li, Xun Lin, Yongli Chu, Wenting Wang, Shiyun Huang, Wei Li, Jin Peng, Chuyao Liu, Wenbin Deng, Chengming Sun, Guanjun Deng.

  The success of tumor immunotherapy highlights the potential of harnessing immune system to fight cancer. Activating both native T cells and exhausted T cells is a critical step for generating effective antitumor immunity, which is determined based on the efficient presentation of tumor antigens and costimulatory signals by antigen-presenting cells, as well as immunosuppressive reversal. However, strategies for achieving an efficient antigen presentation process and improving immunosuppressive microenvironment remain unresolved. Here, aggregation-induced-emission (AIE) photosensitizer-loaded nano-superartificial dendritic cells (saDC@Fs-NPs) were developed by coating superartificial dendritic cells membranes from genetically engineered 4T1 tumor cells onto nanoaggregates of AIE photosensitizers. The outer cell membranes of saDC@Fs-NPs were derived from recombinant lentivirus-infected 4T1 tumor cells in which peptide-major histocompatibility complex class I, CD86 and anti-LAG3 antibody were simultaneously anchored. These saDC@Fs-NPs could directly stimulate T-cell activation and reverse T-cell exhaustion for cancer immunotherapy. The inner AIE-active photosensitizers induced immunogenic cell death to activate dendritic cells and enhance T lymphocyte infiltration by photodynamic therapy, promoting the transformation of "cold tumors" into "hot tumors", which further boosted immunotherapy efficiency. This work presents a powerful photoactive and artificial antigen-presenting platform for activating both native T cells and exhausted T cells, as well as facilitating tumor photodynamic immunotherapy. This article is protected by copyright. All rights reserved.

Keywords:  AIE-active photosensitizers; anti-LAG3 antibody; immunogenic cell death; immunotherapy; superartificial dendritic cells

DOI:  https://doi.org/10.1002/adma.202208555
Nano Lett. 2022 Oct 19.

Probiotic Spore-Based Oral Drug Delivery System for Enhancing Pancreatic Cancer Chemotherapy by Gut-Pancreas-Axis-Guided Delivery.

Zi-Yi Han, Qi-Wen Chen, Zhuang-Jiong Fu, Si-Xue Cheng, Xian-Zheng Zhang.

  The chemotherapeutic effectiveness of pancreatic ductal adenocarcinoma (PDAC) is severely hampered by insufficient intratumoral delivery of antitumor drugs. Here, we demonstrate that enhanced pancreatic cancer chemotherapy can be achieved by probiotic spore-based oral drug delivery system via gut-pancreas axis translocation. Clostridium butyricum spores resistant to harsh external stress are extracted as drug carriers, which are further covalently conjugated with gemcitabine-loaded mesoporous silicon nanoparticles (MGEM). The spore-based oral drug delivery system (SPORE-MGEM) migrates upstream into pancreatic tumors from the gut, which increases intratumoral drug accumulation by ∼3-fold compared with MGEM. In two orthotopic PDAC mice models, tumor growth is markedly suppressed by SPORE-MGEM without obvious side effects. Leveraging the biological contact of the gut-pancreas axis, this probiotic spore-based oral drug delivery system reveals a new avenue for enhancing PDAC chemotherapy.

Keywords:  PDAC chemotherapy; bacterial translocation; gut−pancreas axis; spores; tumor targeting

DOI:  https://doi.org/10.1021/acs.nanolett.2c03131
ACS Nano. 2022 Oct 21.

Enzyme-Catalyzed One-Step Synthesis of Ionizable Cationic Lipids for Lipid Nanoparticle-Based mRNA COVID-19 Vaccines.

Zhongyu Li, Xue-Qing Zhang, William Ho, Fengqiao Li, Mingzhu Gao, Xin Bai, Xiaoyang Xu.

  Ionizable cationic lipid-containing lipid nanoparticles (LNPs) are the most clinically advanced non-viral gene delivery platforms, holding great potential for gene therapeutics. This is exemplified by the two COVID-19 vaccines employing mRNA-LNP technology from Pfizer/BioNTech and Moderna. Herein, we develop a chemical library of ionizable cationic lipids through a one-step chemical-biological enzyme-catalyzed esterification method, and the synthesized ionizable lipids were further prepared to be LNPs for mRNA delivery. Through orthogonal design of experiment methodology screening, the top-performing AA3-DLin LNPs show outstanding mRNA delivery efficacy and long-term storage capability. Furthermore, the AA3-DLin LNP COVID-19 vaccines encapsulating SARS-CoV-2 spike mRNAs successfully induced strong immunogenicity in a BALB/c mouse model demonstrated by the antibody titers, virus challenge, and T cell immune response studies. The developed AA3-DLin LNPs are an excellent mRNA delivery platform, and this study provides an overall perspective of the ionizable cationic lipids, from aspects of lipid design, synthesis, screening, optimization, fabrication, characterization, and application.

Keywords:  COVID-19 vaccines; gene delivery; ionizable lipids; lipid nanoparticle; mRNA therapeutics

DOI:  https://doi.org/10.1021/acsnano.2c07822
Cell. 2022 Oct 11. pii: S0092-8674(22)01196-5. [Epub ahead of print]

Nociceptor neurons direct goblet cells via a CGRP-RAMP1 axis to drive mucus production and gut barrier protection.

Daping Yang, Amanda Jacobson, Kimberly A Meerschaert, Joseph Joy Sifakis, Meng Wu, Xi Chen, Tiandi Yang, Youlian Zhou, Praju Vikas Anekal, Rachel A Rucker, Deepika Sharma, Alexandra Sontheimer-Phelps, Glendon S Wu, Liwen Deng, Michael D Anderson, Samantha Choi, Dylan Neel, Nicole Lee, Dennis L Kasper, Bana Jabri, Jun R Huh, Malin Johansson, Jay R Thiagarajah, Samantha J Riesenfeld, Isaac M Chiu.

  Neuroepithelial crosstalk is critical for gut physiology. However, the mechanisms by which sensory neurons communicate with epithelial cells to mediate gut barrier protection at homeostasis and during inflammation are not well understood. Here, we find that Nav1.8+CGRP+ nociceptor neurons are juxtaposed with and signal to intestinal goblet cells to drive mucus secretion and gut protection. Nociceptor ablation led to decreased mucus thickness and dysbiosis, while chemogenetic nociceptor activation or capsaicin treatment induced mucus growth. Mouse and human goblet cells expressed Ramp1, receptor for the neuropeptide CGRP. Nociceptors signal via the CGRP-Ramp1 pathway to induce rapid goblet cell emptying and mucus secretion. Notably, commensal microbes activated nociceptors to control homeostatic CGRP release. In the absence of nociceptors or epithelial Ramp1, mice showed increased epithelial stress and susceptibility to colitis. Conversely, CGRP administration protected nociceptor-ablated mice against colitis. Our findings demonstrate a neuron-goblet cell axis that orchestrates gut mucosal barrier protection.

Keywords:  CGRP; Ramp1; colitis; goblet cell; gut barrier; gut-brain axis; mucus; neuroepithelial crosstalk; nociceptor; sensory neuron

DOI:  https://doi.org/10.1016/j.cell.2022.09.024
Acta Biomater. 2022 Oct 12. pii: S1742-7061(22)00665-1. [Epub ahead of print]

3D microengineered vascularized tumor spheroids for drug delivery and efficacy testing.

Jungho Ahn, Da-Hyun Kim, Dong-Jun Koo, Jungeun Lim, Tae-Eun Park, Jungseub Lee, Jihoon Ko, Seongchan Kim, Minjae Kim, Kyung-Sun Kang, Dal-Hee Min, Sung-Yon Kim, YongTae Kim, Noo Li Jeon.

  Tumor angiogenesis is regarded as a promising target for limiting cancer progression because tumor-associated vasculature supplies blood and provides a path for metastasis. Thus, in vitro recapitulation of vascularized tumors is critical to understand the pathology of cancer and identify the mechanisms by which tumor cells proliferate, metastasize, and respond to drugs. In this study, we microengineered a vascularized tumor spheroid (VTS) model to reproduce the pathological features of solid tumors. We first generated tumor-EC hybrid spheroids with self-assembled intratumoral vessels, which enhanced the uniformity of the spheroids and peritumoral angiogenic capacity compared to spheroids composed only with cancer cells. Notably, the hybrid spheroids also exhibited expression profiles associated with aggressive behavior. The blood vessels sprouting around the hybrid spheroids on the VTS chip displayed the distinctive characteristics of leaky tumor vessels. With the VTS chip showing a progressive tumor phenotype, we validated the suppressive effects of axitinib on tumor growth and angiogenesis, which depended on exposure dose and time, highlighting the significance of tumor vascularization to predict the efficacy of anticancer drugs. Ultimately, we effectively induced both lymphangiogenesis and angiogenesis around the tumor spheroid by promoting interstitial flow. Thus, our VTS model is a valuable platform with which to investigate the interactions between tumor microenvironments and explore therapeutic strategies in cancer. STATEMENT OF SIGNIFICANCE: : We conducted an integrative study within a vascularized tumor spheroid (VTS) model. We first generated tumor-EC hybrid spheroids with self-assembled intratumoral vessels, which enhanced the uniformity of the spheroids and peritumoral angiogenic capacity compared to spheroids composed only with cancer cells. Through RNA sequencing, we elucidated that the tumor-EC hybrid spheroids exhibited expression profiles associated with aggressive behavior such as cancer progression, invasion and metastasis. The blood vessels sprouting around the hybrid spheroids on the VTS chip displayed the distinctive characteristics of leaky tumor vessels. We further validated the suppressive effects of axitinib on tumor growth and angiogenesis, which depended on exposure dose and time. Ultimately, we effectively induced both lymphangiogenesis and angiogenesis around the tumor spheroid by promoting interstitial flow.

Keywords:  Drug screening; Leaky tumor vasculature; Microfluidics; Organ-on-a-Chip; Tumor angiogenesis; Tumor microenvironment; Vascularized tumor

DOI:  https://doi.org/10.1016/j.actbio.2022.10.009
Adv Mater. 2022 Oct 17. e2205794

Drug Delivery in Plants Using Silk Microneedles.

Yunteng Cao, Sally Shuxian Koh, Yangyang Han, Javier Jingheng Tan, Doyoon Kim, Nam-Hai Chua, Daisuke Urano, Benedetto Marelli.

  New systems for agrochemical delivery in plants will foster precise agricultural practices and provide new tools to study plants and design crop traits, as standard spray methods suffer from elevated loss and limited access to remote plant tissues. Silk-based microneedles can circumvent these limitations by deploying a known amount of payloads directly in plants' deep tissues. However, plant response to microneedles' application and microneedles' efficacy in deploying physiologically relevant biomolecules are unknown. Here, we show that gene expression associated with Arabidopsis thaliana wounding response decreases within 24 hours post microneedles' application. Additionally, microinjection of gibberellic acid (GA3 ) in A. thaliana mutant ft-10 provides a more effective and efficient mean than spray to activate GA3 pathways, accelerating bolting, and inhibiting flower formation. Microneedles' efficacy in delivering GA3 is also observed in several monocot and dicot crop species, i.e., tomato (Solanum lycopersicum), lettuce (Lactuca sativa), spinach (Spinacia oleracea), rice (Oryza Sativa), maize (Zea mays), barley (Hordeum vulgare), and soybean (Glycine max). The wide range of plants that can be successfully targeted with microinjectors opens the doors to their use in plant science and agriculture. This article is protected by copyright. All rights reserved.

Keywords:  gibberellins; payload delivery; plants’ wounding responses; silk microneedles; sustainable agriculture

DOI:  https://doi.org/10.1002/adma.202205794
Proc Natl Acad Sci U S A. 2022 Oct 25. 119(43): e2213373119

Photoacoustic image-guided biomimetic nanoparticles targeting rheumatoid arthritis.

Jianhai Chen, Silue Zeng, Qiang Xue, Yuchuan Hong, Liangjian Liu, Liang Song, Chihua Fang, Hai Zhang, Benguo Wang, Adam C Sedgwick, Peng Zhang, Jonathan L Sessler, Chengbo Liu, Jingqin Chen.

  The high level of reactive oxygen species (ROS) in the rheumatoid arthritis (RA) microenvironment (RAM) and its persistent inflammatory nature can promote damage to joints, bones, and the synovium. Targeting strategies that integrate effective RAM regulation with imaging-based monitoring could lead to improvements in the diagnosis and treatment of RA. Here, we report the combined use of small interfering RNAs (siRNAsT/I) and Prussian blue nanoparticles (PBNPs) to silence the expression of proinflammatory cytokines TNF-α/IL-6 and scavenge the ROS associated with RAM. To enhance the in vitro and in vivo biological stability, biocompatibility, and targeting capability of the siRNAsT/I and PBNPs, macrophage membrane vesicles were used to prepare biomimetic nanoparticles, M@P-siRNAsT/I. The resulting constructs were found to suppress tumor necrosis factor-α/interleukin-6 expression and overcome the hypoxic nature of RAM, thus alleviating RA-induced joint damage in a mouse model. The M@P-siRNAsT/I of this study could be monitored via near-infrared photoacoustic (PA) imaging. Moreover, multispectral PA imaging without the need for labeling permitted the real-time evaluation of M@P-siRNAsT/I as a putative RA treatment. Clinical microcomputed tomography and histological analysis confirmed the effectiveness of the treatment. We thus suggest that macrophage-biomimetic M@P-siRNAsT/I and their analogs assisted by PA imaging could provide a new strategy for RA diagnosis, treatment, and monitoring.

Keywords:  biomimetic nanoparticles; photoacoustic; rheumatoid arthritis; small interfering RNA; targeted imaging

DOI:  https://doi.org/10.1073/pnas.2213373119
Nat Commun. 2022 Oct 19. 13(1): 6203

Remodelling of tumour microenvironment by microwave ablation potentiates immunotherapy of AXL-specific CAR T cells against non-small cell lung cancer.

Bihui Cao, Manting Liu, Lu Wang, Kangshun Zhu, Mingyue Cai, Xiaopei Chen, Yunfei Feng, Shuo Yang, Shengyu Fu, Cheng Zhi, Xiaodie Ye, Jian Zhang, Zhiru Zhang, Xin Yang, Ming Zhao, Qingde Wu, Linfeng Xu, Lili Yang, Hui Lian, Qi Zhao, Zhenfeng Zhang.

The complex immunosuppressive tumour microenvironment (TME) and lack of tumour-specific targets hinder the application of chimeric antigen receptor (CAR) T cells in the treatment of solid tumours. Combining local treatment with CAR T cell immunotherapy may regulate the TME and enhance the killing potency of CAR T cells in solid tumours. Here, we show that AXL, which is highly expressed in non-small cell lung cancer (NSCLC) but not in normal tissues, might be a target for CAR T cell therapy. AXL-CAR T cells alone cause moderate tumour regression in subcutaneous and pulmonary metastatic lung cancer cell-derived xenograft models. Combination of microwave ablation (MWA) and AXL-CAR T cells have superior antitumour efficacy. MWA enhances the activation, infiltration, persistence and tumour suppressive properties of AXL-CAR T cells in AXL-positive NSCLC patient-derived xenograft tumours via TME remodelling. The combination therapy increases the mitochondrial oxidative metabolism of tumour-infiltrating CAR T cells. Combination treatment induces significant tumour suppression without observed toxicities in humanized immunocompetent mice. The synergistic therapeutic effect of MWA and AXL-CAR T cells may be valuable for NSCLC treatment.

DOI: https://doi.org/10.1038/s41467-022-33968-5
Nanomedicine. 2022 Oct 15. pii: S1549-9634(22)00096-X. [Epub ahead of print] 102610

A combinational chemo-immune therapy using outer membrane vesicles for enhanced cancer therapy by RGD targeting.

Shuping Li, Xiaodong Gao.

  Cancer therapies are limited by poor drug penetration that impedes effective tumor treatment. This was overcome in the present study by loading the immune reaction inducing nanocarriers of the bacterial outer membrane vesicles (OMVs) and doxorubicin (DOX) into the natural immunity platform OMV via incubation. Drug accumulation at the tumor site was improved by using the targeting peptide 6-Mal- Arg-Gly-Asp (RGD) on the surface of OMVs to increase internalization via binding to cell surface integrin αvβ3. OMVs stimulate immune responses by reversing the immune-suppressive tumor microenvironment (TME) via decreasing TAM and Treg, increasing CD8+ T and M1, and promoting DC maturation. The combination of DOX and OMVs compensates for the shortcomings of monotherapy (e.g., chemotherapy and immunotherapy) and amplifies the therapeutic efficacy of cancer treatment, while aiding selection of novel nanocarriers and development of effective therapeutic regimens.

Keywords:  DC maturation; DOX; OMV; RGD; TME

DOI:  https://doi.org/10.1016/j.nano.2022.102610
ACS Nano. 2022 Oct 18.

Triggered Self-Sorting of Peptides to Form Higher-Order Assemblies in a Living System.

Xuejiao Yang, Honglei Lu, Bihan Wu, Huaimin Wang.

  Biological components (protein, DNA, lipid rafts, etc.) self-sort to form higher-order structures with elegant modulation by endogenous stimuli for maintaining cellular functions in living cells. However, the challenge of producing self-sorted higher-order assemblies of peptides in living systems (cells and tissues) spatiotemporally has yet to be achieved. This work reports the using of a biocompatible strategy to construct self-sorted assemblies of peptides in living cells and tumor-bearing mice. The results show that the designed peptides self-sort to form distinct nanostructures in living cancer cells using an endogenous trigger, as evidenced by confocal laser scanning microscopy and Bio-EM. Wound-healing experiments indicate that the in situ generation of self-sorted nanostructures exhibits a synergistic effect that significantly decreases the migration of cancer cells. In vivo experiments demonstrate that the designed peptides could self-sort in tumor-bearing mice and improve the tumor penetrating ability of the impenetrable component in tumor tissue. We can further program the formation of self-sorted materials through orthogonal triggers by introducing an exogenous trigger (light) and an endogenous trigger independently. Thus, this work provides a strategy to control multiple self-assembling processes in the context of the living system and provides a general strategy to construct self-sorted structures for the emergent properties of materials science.

Keywords:  higher-order assemblies; nanofibers; nanospheres; peptide; self-sorting

DOI:  https://doi.org/10.1021/acsnano.2c05825
Adv Mater. 2022 Oct 21. e2205498

Photoswitchable Microgel for Dynamic Macrophage Modulation.

Yuri Kim, Ramar Thangam, Jounghyun Yoo, Jeongyun Heo, Jung Yeon Park, Nayeon Kang, Sungkyu Lee, Jiwon Yoon, Kwang Rok Mun, Misun Kang, Sunhong Min, Seong Yeol Kim, Subin Son, Jihwan Kim, Hyunsik Hong, Gunhyu Bae, Kanghyeon Kim, Sanghyeok Lee, Letao Yang, Ja Yeon Lee, Jinjoo Kim, Steve Park, Dong-Hyun Kim, Ki-Bum Lee, Woo Young Jang, Bong Hoon Kim, Ramasamy Paulmurugan, Seung-Woo Cho, Hyun-Cheol Song, Seok Ju Kang, Wujin Sun, Yangzhi Zhu, Junmin Lee, Han-Jun Kim, Ho Seong Jang, Jong Seung Kim, Ali Khademhosseini, Yongju Kim, Sehoon Kim, Heemin Kang.

  Dynamic manipulation of supramolecular self-assembled structures has been achieved irreversibly or under non-physiological conditions, thereby limiting their biomedical, environmental, and catalysis applicability. In this study, microgels composed of azobenzene derivatives stacked via π-cation and π-π interactions are developed that are electrostatically stabilized with Arg-Gly-Asp (RGD)-bearing anionic polymers. Lateral swelling of RGD-bearing microgels occurs via cis-azobenzene formation mediated by near-infrared light-upconverted-ultraviolet light, which disrupts intermolecular interactions on the visible light-absorbing upconversion nanoparticle-coated materials. Real-time imaging and molecular dynamics simulations demonstrate the deswelling of RGD-bearing microgels via visible light-mediated trans-azobenzene formation. Near-infrared light can induce in situ swelling of RGD-bearing microgels to increase RGD availability and trigger release of loaded interleukin-4 that facilitates the adhesion structure assembly linked with pro-regenerative polarization of host macrophages. In contrast, visible light can induce deswelling of RGD-bearing microgels to decrease RGD availability that suppresses macrophage adhesion that yields pro-inflammatory polarization. Our microgels exhibits high stability and non-toxicity. Versatile use of ligands and protein delivery can offer cytocompatible and photoswitchable manipulability of diverse host cells. This article is protected by copyright. All rights reserved.

Keywords:  Photoswitchable microgel; dynamic hydrogel; macrophage adhesion; macrophage polarization; microgel swelling

DOI:  https://doi.org/10.1002/adma.202205498
Eur J Pharm Biopharm. 2022 Oct 17. pii: S0939-6411(22)00238-7. [Epub ahead of print]

Lipidoid nanoparticles increase ATP uptake into hypoxic brain endothelial cells.

Purva Khare, James F Conway, Devika S Manickam.

  Lipidoid nanoparticles (LNPs) are clinically successful carriers for nucleic acid delivery to liver and muscle targets. The ability of LNPs to load and deliver small molecule drugs has not been reported yet. We propose that the delivery of adenosine triphosphate (ATP) to brain endothelial cells (BECs) lining the blood-brain barrier may increase cellular energetics of the injured BECs. We formulated and studied the physicochemical characteristics of ATP-loaded LNPs using the C12-200 ionizable cationic lipid and other helper lipids. Polyethylene glycol-dimyristoyl glycerol (PEG-DMG), one of the helper lipids, played a crucial role in maintaining colloidal stability of LNPs over time whereas the inclusion of both ATP and PEG-DMG maintained the colloidal stability of LNPs in the presence of serum proteins. ATP-LNPs formulated with PEG-DMG resulted in a 7.7- and 6.6- fold increased uptake of ATP into normoxic and hypoxic BECs, respectively. Altogether, our results demonstrate the potential of LNPs as a novel carrier for the delivery of small molecular mass actives to BECs-a CNS target.

Keywords:  ATP delivery; C12-200; Lipidoid nanoparticle; PEG-DMG; brain endothelial cells

DOI:  https://doi.org/10.1016/j.ejpb.2022.10.011
Nat Commun. 2022 Oct 21. 13(1): 6279

Lactose azocalixarene drug delivery system for the treatment of multidrug-resistant pseudomonas aeruginosa infected diabetic ulcer.

Juan-Juan Li, Yuqing Hu, Bing Hu, Wenbo Wang, Haiqi Xu, Xin-Yue Hu, Fei Ding, Hua-Bin Li, Ke-Rang Wang, Xinge Zhang, Dong-Sheng Guo.

Diabetic wound is one of the most intractable chronic wounds that is prone to bacterial infection. Hypoxia is an important feature in its microenvironment. However, it is challenging for antimicrobial therapy to directly apply the existing hypoxia-responsive drug delivery systems due to the active targeting deficiency and the biofilm obstacle. Herein, we customizes a hypoxia-responsive carrier, lactose-modified azocalix[4]arene (LacAC4A) with the ability to actively target and inhibit biofilm. By loading ciprofloxacin (Cip), the resultant supramolecular nanoformulation Cip@LacAC4A demonstrates enhanced antibacterial efficacy resulting from both the increased drug accumulation and the controlled release at the site of infection. When applied on diabetic wounds together with multidrug-resistant Pseudomonas aeruginosa infection in vivo, Cip@LacAC4A induces definitely less inflammatory infiltration than free Cip, which translates into high wound healing performance. Importantly, such design principle provides a direction for developing antimicrobial drug delivery systems.

DOI: https://doi.org/10.1038/s41467-022-33920-7
Adv Sci (Weinh). 2022 Oct 21. e2201988

Machine Learning Guides Peptide Nucleic Acid Flow Synthesis and Sequence Design.

Chengxi Li, Genwei Zhang, Somesh Mohapatra, Alex J Callahan, Andrei Loas, Rafael Gómez-Bombarelli, Bradley L Pentelute.

  Peptide nucleic acids (PNAs) are potential antisense therapies for genetic, acquired, and viral diseases. Efficiently selecting candidate PNA sequences for synthesis and evaluation from a genome containing hundreds to thousands of options can be challenging. To facilitate this process, this work leverages machine learning (ML) algorithms and automated synthesis technology to predict PNA synthesis efficiency and guide rational PNA sequence design. The training data is collected from individual fluorenylmethyloxycarbonyl (Fmoc) deprotection reactions performed on a fully automated PNA synthesizer. The optimized ML model allows for 93% prediction accuracy and 0.97 Pearson's r. The predicted synthesis scores are validated to be correlated with the experimental high-performance liquid chromatography (HPLC) crude purities (correlation coefficient R2 = 0.95). Furthermore, a general applicability of ML is demonstrated through designing synthetically accessible antisense PNA sequences from 102 315 predicted candidates targeting exon 44 of the human dystrophin gene, SARS-CoV-2, HIV, as well as selected genes associated with cardiovascular diseases, type II diabetes, and various cancers. Collectively, ML provides an accurate prediction of PNA synthesis quality and serves as a useful computational tool for informing PNA sequence design.

Keywords:  automated synthesis; drug design; machine learning; peptide nucleic acid; yield prediction

DOI:  https://doi.org/10.1002/advs.202201988
J Control Release. 2022 Oct 17. pii: S0168-3659(22)00690-3. [Epub ahead of print]

Airway epithelial cell-specific delivery of lipid nanoparticles loading siRNA for asthma treatment.

Mengjun Zhang, Huiyang Jiang, Lan Wu, Haoyu Lu, Hriday Bera, Xing Zhao, Xiong Guo, Xulu Liu, Dongmei Cun, Mingshi Yang.

  With specific and inherent mRNA cleaving activity, small interfering RNA (siRNA) has been deemed promising therapeutics to reduce the exacerbation rate of asthma by inhibiting the expression and release of proinflammatory cytokines from airway epithelial cells (AECs). To exert the therapeutic effects of siRNA drugs, nano-formulations with high efficiency and safety are required to deliver these nucleic acids to the target cells. Herein, we exploited novel inhaled lipid nanoparticles (LNPs) targeting intercellular adhesion molecule-1 (ICAM-1) receptors on the apical side of AECs. This delivery system is meant to enhance the specific delivery efficiency of siRNA in AECs to prevent the expression of proinflammatory cytokines in AECs and the concomitant symptoms in parallel. A cyclic peptide that resembles part of the capsid protein of rhinovirus and binds to ICAM-1 receptors was initially conjugated with cholesterol and subsequently assembled with ionizable cationic lipids to form the LNPs (Pep-LNPs) loaded with siRNA against thymic stromal lymphopoietin (TSLP siRNA). The obtained Pep-LNPs were subjected to thorough characterization and evaluations in vitro and in vivo. Pep-LNPs significantly enhanced cellular uptake and gene silencing efficiency in human epithelial cells expressing ICAM-1 in vitro, exhibited AEC-specific delivery and improved the gene silencing effect in ovalbumin-challenged asthmatic mice after pulmonary administration. More importantly, Pep-LNPs remarkably downregulated the expression of TSLP in AECs, effectively alleviated inflammatory cell infiltration, and reduced the secretion of other proinflammatory cytokines, including IL-4 and IL-13, as well as mucus production in asthmatic mice. This study demonstrates that Pep-LNPs are safe and efficient to deliver siRNA drugs to asthmatic AECs and could potentially alleviate allergic asthma by inhibiting the overexpression of proinflammatory cytokines in the airway.

Keywords:  Airway epithelial cells; Asthma; Lipid nanoparticles; Pulmonary delivery; siRNA

DOI:  https://doi.org/10.1016/j.jconrel.2022.10.020
Proc Natl Acad Sci U S A. 2022 Oct 25. 119(43): e2211065119

Targeted delivery of an anti-inflammatory corticosteroid to Ly6C/G-positive cells abates severity of influenza A symptoms.

Novalia Pishesha, Thibault Harmand, Claire Carpenet, Xin Liu, Atul Bhan, Ashraful Islam, Renate van den Doel, William Pinney, Hidde L Ploegh.

  The distribution of Ly6C/G-positive cells in response to an infection of the mouse respiratory tract with influenza A virus was followed noninvasively over time by immuno-positron emission tomography. We converted nanobodies that recognize Ly6C and Ly6G, markers of neutrophils and other myeloid cells, as well as an influenza hemagglutinin-specific nanobody, into 89Zr-labeled PEGylated positron emission tomography (PET) imaging agents. The PET images showed strong accumulation of these imaging agents in the lungs of infected mice. Immunohistochemistry of influenza virus-infected mice and control mice, injected with a biotinylated and PEGylated version of the Ly6C/G-specific nanobody, showed the presence of abundant Ly6C/G-positive myeloid cells and positivity for Ly6C/G on bronchial epithelium in influenza virus-infected mice. This is consistent with focal inflammation in the lungs, a finding that correlated well with the immuno-PET results. No such signals were detected in control mice. Having shown by PET the accumulation of the Ly6C/G-specific nanobody in infected lungs, we synthesized conjugates of Ly6C/G-specific nanobodies with dexamethasone to enable targeted delivery of this immunosuppressive corticosteroid to sites of inflammation. Such conjugates reduced the weight loss that accompanies infection, while the equivalent amount of free dexamethasone was without effect. Nanobody-drug conjugates thus enable delivery of drugs to particular cell types at the appropriate anatomic site(s). By avoiding systemic exposure to free dexamethasone, this strategy minimizes its undesirable side effects because of the much lower effective dose of the nanobody-dexamethasone conjugate. The ability to selectively target inflammatory cells may find application in the treatment of other infections or other immune-mediated diseases.

Keywords:  PET imaging; drug conjugates; inflammation; nanobody; neutrophils

DOI:  https://doi.org/10.1073/pnas.2211065119