Efficient Delivery of dsRNA and DNA in Cultured Silkworm Cells for Gene Function Analysis Using PAMAM Dendrimers System

Lü, Chenchen; Li, Zhiqing; Chang, Li; Dong, Zhaoming; Guo, Pengchao; Shen, Guanghui; Xia, Qingyou

doi:10.3390/insects11010012

Cited by 13 publications

(6 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The molecular weight of NDX is approximately 20MDa and poly I:C is approximately 2kDa, therefore at this loading ratio there are approximately 10000 molecules of poly I:C per molecule of NDX (NCBI, 2022; Besford et al, 2020). To compare to other nanoparticles, generation 5 poly(amidoamine) (G5-PAMAM) bound a 250bp in vitro transcribed molecule at a nanoparticle:dsRNA ratio of 3:2, while PAMAM dendrimer-coated magnetic nanoparticles required a ratio of 11:1 or greater (Lu et al, 2019;Khodadust et al, 2013). Ferumoxytol, an approved iron oxide nanoparticle, complete poly I:C binding was lost at the w/w ratio 10:1 (Zhao et al, 2018).…”

Section: Resultsmentioning

confidence: 99%

Phytoglycogen-dsRNA nanoparticles demonstrate differential cytotoxicity and immunostimulatory potential in two ovarian cancer cell lines

Lewis

Tran

Aldor

et al. 2022

Preprint

View full text Add to dashboard Cite

Background Ovarian cancer is a leading cause of cancer mortality in women, and only a small percentage of cases are caught at an early stage. Novel treatments with improved efficacy are needed to fight ovarian cancer and to overcome resistance to traditional therapies. Double-stranded (ds) RNA, including the synthetic polyinosinic cytidylic acid (poly (I:C), has shown promise as a cancer therapeutic. Two ovarian cancer cell lines were tested for their ability to produce an immune response to poly (I:C) delivered using a nanoparticle carrier, a biodegradable phytoglycogen derived from sweet corn, called nanodendrix (NDX). SKOV-3 and OVCAR-3 have been previously identified as dsRNA-resistant and dsRNA-sensitive, respectively. Results Firstly, NDX was found to effectively bind poly (I:C), at a w/w ratio of 2:1 NDX:poly (I:C), the resulting particles, poly (I:C)-NDX, were tested for biological activity through uptake and two therapeutic modes of action, cytotoxicity and stimulation of the innate immune response. Both cell lines bound poly (I:C)-NDX, as observed using immunocytochemistry. In OVCAR-3 poly (I:C)-NDX caused significant cell death, even at concentrations as low as 62.5ng/mL, measured using the cell viability indicator dye alamarBlue; no cell death was observed with poly (I:C) alone across all concentrations, up to 5µg/mL in SKOV-3 and 0.5µg/mL in OVCAR-3. In both OVCAR-3 and SKOV-3, poly (I:C)-NDX stimulated the production of an innate immune chemokine, CXCL10, at the transcript and protein levels, at significantly higher levels than poly (I:C) alone. Interestingly, in response to poly (I:C)-NDX SKOV-3 produced a more robust immune response compared and higher levels of capase-3/-7 activation compared to OVCAR-3, despite showing no significant cell death. Conclusions Poly (I:C)-NDX represents a robust and multifunctional therapy with demonstrated efficacy against a range of ovarian tumour cells, potentiating poly (I:C) and sensitizing resistant cells. Additionally, the SKOV-3 and OVCAR-3 combination represents a powerful comparative model to help unravel dsRNA-mediated immune responses in ovarian cancer cells.

show abstract

Section: Resultsmentioning

confidence: 99%

Phytoglycogen-dsRNA nanoparticles demonstrate differential cytotoxicity and immunostimulatory potential in two ovarian cancer cell lines

Lewis

Tran

Aldor

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…[ 146 ] Currently, the ability of vectors such as PAMAM dendrimers to deliver RNA to tumor cells has been well established. [ 147 ] Multiple in vivo and in vitro studies have proved that PAMAM dendrimers can successfully deliver genetic material (siRNA, miRNA, and plasmids) into cells and produce specific therapeutic effects, [ 130 ] such as silencing some protein genes and inhibiting the proliferation of tumor cells.…”

Section: Utilization Of Dendrimers In Cancermentioning

confidence: 99%

Polymeric Dendrimers as Nanocarrier Vectors for Neurotheranostics

Zha

Liu

et al. 2022

Small

View full text Add to dashboard Cite

immune responses, activating cellular machinery causing cell stress, cytotoxicity, lack of selectivity, increasing the likelihood of replication competence, and the constrained structure of insert-size, which can limit their biological and medical applications. [2][3][4] By contrast, various forms of nonviral vectors are widely developed and applied in biomedical research because of their low immunogenicity, high biocompatibility, simplicity in structural pattern, ease of size adjustment, practicality in designing various active sites, and low cost of production. [5,6] The most widely studied nonviral vectors are liposomes, inorganic nanoparticles, polymers, and dendrimers. [7] Liposomes are small spherical vesicles from cholesterol and phospholipids characterized with the surface that can be modified using glycolipids or sialic acid. Yet, the unmodified forms of liposomes are structurally unstable and could easily be eliminated by means of body circulation. [8] In this situation, stable inorganic nanoparticles, such as calcium phosphate (Ca 3 (PO 4 ) 2 ), gold (Au), iron oxide (Fe 3 O 4 ), and lanthanide-based nanoparticles are widely used, mainly due to their good hydrophilicity and excellent biocompatibility. [9][10][11] Depending on their chemical core composition, these inorganic nanoparticles display different properties and efficiencies. Lately, inorganic nanoparticles have been developed as effective drug delivery systems and used in many diagnostic techniques. [12] In addition, polymers are widely used as nonviral vectors as well. [13][14][15] Polymers are biodegradable nanomaterials with stable and easily modifiable 3D structures. Their uncomplicated synthesis process allows for large-scale production and safe clinical use compared to inorganic nanoparticles. [16] Notably, dendrimers are hyperbranched polymers with a well-defined chemical structure: a core at the center occupied by array of convergent reactive chain-ends and surface that can easily be modified. [17] The chemistry of dendrimers was first described by Buhleier et al. in 1978. [18] In 1985, Tomalia et al. reported a newer kind of dendrimers based on the mixture of amines and amides, termed polyamidoamine (PAMAM) dendrimers. [19] Dendrimers are intermeshed synthetic organic macromolecules with 3D architecture composition and abundant terminal functional groups. They are well-defined monodispersity-sized materials with extraordinary membrane interaction properties Dendrimers are polymers with well-defined 3D branched structures that are vastly utilized in various neurotheranostics and biomedical applications, particularly as nanocarrier vectors. Imaging agents can be loaded into dendrimers to improve the accuracy of diagnostic imaging processes. Likewise, combining pharmaceutical agents and anticancer drugs with dendrimers can enhance their solubility, biocompatibility, and efficiency. Practically, by modifying ligands on the surface of dendrimers, effective therapeutic and diagnostic platforms can be constructed and implemented for target...

show abstract

“…The dsRNAs for control gene of Red were also synthesized. G5-PAMAMmediated delivery of dsRNA was used according to the previous protocol (45). G5-PAMAM was purchased from Chenyuan Company (Shandong, China).…”

Section: Rna Interferencementioning

confidence: 99%

“…We firstly synthesized specific dsRNAs against ORF and UTR of BmTCTP (referred to as dsTCTP-ORF and dsTCTP-UTR) (Figure 2A). G5-PAMAM system was used to deliver dsRNAs of BmTCTP into the BmN cells to silence BmTCTP expression (45). RT-PCR result showed that both dsTCTP-ORF and dsTCTP-UTR were able to decrease the mRNA transcription of BmTCTP compared with the dsRed control treatment (Figure 2B), and also dsTCTP-ORF could specifically decrease the expression of FLAG-TCTP by immunoblotting (Figure S1), which together revealed that dsRNAs of BmTCTP could efficiently silence its own expression.…”

Section: Bmtctp Knockdown Promoted Viral Replicationmentioning

confidence: 99%

SUMOylation of Translationally Regulated Tumor Protein Modulates Its Immune Function

Lü

Zhang

et al. 2022

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

Translationally controlled tumor protein (TCTP) is a highly conserved protein possessing numerous biological functions and molecular interactions, ranging from cell growth to immune responses. However, the molecular mechanism by which TCTP regulates immune function is largely unknown. Here, we found that knockdown of Bombyx mori translationally controlled tumor protein (BmTCTP) led to the increased susceptibility of silkworm cells to virus infection, whereas overexpression of BmTCTP significantly decreased the virus replication. We further demonstrated that BmTCTP could be modified by SUMOylation molecular BmSMT3 at the lysine 164 via the conjugating enzyme BmUBC9, and the stable SUMOylation of BmTCTP by expressing BmTCTP-BmSMT3 fusion protein exhibited strong antiviral activity, which confirmed that the SUMOylation of BmTCTP would contribute to its immune responses. Further work indicated that BmTCTP is able to physically interact with interleukin enhancer binding factor (ILF), one immune molecular, involved in antivirus, and also induce the expression of BmILF in response to virus infection, which in turn enhanced antiviral activity of BmTCTP. Altogether, our present study has provided a novel insight into defending against virus via BmTCTP SUMOylation signaling pathway and interacting with key immune molecular in silkworm.

show abstract

Efficient Delivery of dsRNA and DNA in Cultured Silkworm Cells for Gene Function Analysis Using PAMAM Dendrimers System

Cited by 13 publications

References 34 publications

Phytoglycogen-dsRNA nanoparticles demonstrate differential cytotoxicity and immunostimulatory potential in two ovarian cancer cell lines

Phytoglycogen-dsRNA nanoparticles demonstrate differential cytotoxicity and immunostimulatory potential in two ovarian cancer cell lines

Polymeric Dendrimers as Nanocarrier Vectors for Neurotheranostics

SUMOylation of Translationally Regulated Tumor Protein Modulates Its Immune Function

Contact Info

Product

Resources

About