Dendritic peptide-conjugated polymeric nanovectors for non-toxic delivery of plasmid DNA and enhanced non-viral transfection of immune cells

Yi, Sijia; Kim, Sun‐Young; Vincent, Michael; Yuk, Simseok A; Bobbala, Sharan; Du, Fanfan; Scott, Evan A.

doi:10.1016/j.isci.2022.104555

Cited by 6 publications

(18 citation statements)

References 86 publications

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“…Commercially available cationic lipidbased transfection reagent Lipofectamine TM 2000 and Lipofectamine TM 3000 can transfect many cultured animal cells [53]. However, they are significantly cytotoxic [53] and are not efficient in delivering genes into hard-to-transfect RAW 264.7 cells [54,55]. Notably, liposomes containg only RGDKlipopeptide I and DOPE (in 1:1 mole ratio) were found to be somewhat less efficient (29%) in transfecting RAW 264.7 cells.…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

RGDK-lipopeptide for targeting genetic vaccines to antigen presenting cells

Rahaman,

Chaudhuri

2023

Biomed. Mater.

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To ensure effective immune response in genetic immunizations, DNA/mRNA vaccines need to be delivered to body’s antigen presenting cells (APCs) which is a challenging task. This is primarily due to presence of high concentrations of various degradative enzymes inside them. To this end, mannose receptor (over expressed in APCs) selective cationic liposomes have been used in the past for delivering antigen-encoded plasmid DNA to APCs. APCs also express integrin receptors on their cell surfaces. However, studies aimed at delivering DNA vaccines into APCs via integrin receptors have not yet been undertaken. Herein, we report on the use of cationic liposomes of a priorly disclosed α5β1 integrin receptor selective RGDK-lipopeptide for macrophage transfection. In this study, we have used pCMV-GFP (as model DNA vaccine) and RAW 264.7 cells (mouse macrophages cells) as model APC. We show that the liposomes of RGDK-lipopeptide containing a previously reported endosome-disrupting histidinylated lipid and DOPE (as co-lipid) in 0.5:0.5:1.0 mole ratio are the most competent in transfecting macrophage cells (44%). Findings in the fluorescence resonance energy transfer based membrane fusogencity assay revealed that the enhanced macrophage transfection efficiency of the liposomes containing RGDK-lipopeptide, endosome-disrupting histidinylated and DOPE may originate from its higher membrane fusogenicity than that for liposomes containing only RGDK-lipopeptide and DOPE. The presently described biologically safe liposomal formulations of RGDK-lipopeptide are expected to find biomedical applications in future for combating cancer and infectious diseases through genetic immunizations.

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Section: Cytotoxicity Assaymentioning

confidence: 99%

RGDK-lipopeptide for targeting genetic vaccines to antigen presenting cells

Rahaman,

Chaudhuri

2023

Biomed. Mater.

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“…Cytotoxicity was assessed by staining cells with crystal violet. Briefly, 2 x 10 4 cells/well were cultured in 24-well plates and treated with either 0.25, 0.5, 1, 1.5 or 2 µg rrsp-mRNA by PPDP2 nanocarriers or 2 µg rrsp-mRNA via MessengerMAX lipofectamine (Invitrogen) for 72 h. Cells were washed and crystal violet fixing/staining solution was added for 20 minutes at room temperature as described previously (20). Images of air-dried plates were acquired using a conventional desktop scanner.…”

Section: Cytotoxicity and Apoptosis Assaysmentioning

confidence: 99%

“…no. 600-401-P16] antibodies as described previously (20). Primary antibodies were detected using the appropriate secondary antibodies and 3,3 0 -diaminobenzidine revelation (Dako).…”

Section: Histology Ihc and Image Analysismentioning

confidence: 99%

Therapeutic expression of RAS Degrader RRSP in Pancreatic Cancer via Nanocarrier-mediated mRNA delivery

Escher,

Yuk,

Qian

et al. 2024

Preprint

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FigureGraphical Abstract:Synthetic nanocarriers packaged with mRNA are used to express the RAS-specific protease RRSP in cancer cells resulting in cell death and tumor shrinkage.About one-third of all human cancers encode abnormal RAS proteins locked in a constitutively activated state to drive malignant transformation and uncontrolled tumor growth. Despite progress in development of small molecules for treatment of mutant KRAS cancers, there is a need for a pan-RAS inhibitor that is effective against all RAS isoforms and variants and that avoids drug resistance. We have previously shown that the naturally occurring bacterial enzyme RAS/RAP1-specific endopeptidase (RRSP) is a potent RAS degrader that can be re-engineered as a biologic therapy to induce regression of colorectal, breast, and pancreatic tumors. Here, we have developed a strategy for in vivo expression of this RAS degrader via mRNA delivery using a synthetic nonviral gene delivery platform composed of the poly(ethylene glycol)-b-poly(propylene sulfide) (PEG-b-PPS) block copolymer conjugated to a dendritic cationic peptide (PPDP2). Using this strategy, PPDP2 is shown to deliver mRNA to both human and mouse pancreatic cells resulting in RRSP gene expression, activity, and loss of cell proliferation. Further, pancreatic tumors are reduced with residual tumors lacking detectable RAS and phosphorylated ERK. These data support that mRNA-loaded synthetic nanocarrier delivery of a RAS degrader can interrupt the RAS signaling system within pancreatic cancer cells while avoiding side effects during therapy.

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“…Another example is the poly(ethylene glycol)- b -poly(propylene sulfide)-cationic dendritic peptide copolymer (PEG- b -PPS-ss-DP, PPDP), which is based on reduceable bonding and connected with cationic dendritic peptide fragments. 45 Its dendritic three-dimensional structure and small cationic charge ratio make the polymer not only have the specific advantages of high encapsulation capacity, but also high transfection efficiency and lower toxicity. In this regard, PEG 17 - b -PPS 80 -DP (PPDP2) was encapsulated with a pDNA plasmid in a 60 : 1 ratio, which effectively escaped macrophage lysosomes within 18 hours, promoted the delivery and release of the pDNA plasmid in cytoplasm, and increased the concentration of plasmid accumulation in macrophages (Fig.…”

Section: Stimulus-responsive Polymersmentioning

confidence: 99%