key: cord-273347-eyxc4rt0
authors: Mohammadinejad, Reza; Dehshahri, Ali; Madamsetty, Vijay Sagar; Zahmatkeshan, Masoumeh; Tavakol, Shima; Makvandi, Pooyan; Khorsandi, Danial; Pardakhty, Abbas; Ashrafizadeh, Milad; Afshar, Elham Ghasemipour; Zarrabi, Ali
title: In vivo gene delivery mediated by non-viral vectors for cancer therapy
date: 2020-07-04
journal: J Control Release
DOI: 10.1016/j.jconrel.2020.06.038
sha: 
doc_id: 273347
cord_uid: eyxc4rt0

Gene therapy by expression constructs or down-regulation of certain genes has shown great potential for the treatment of various diseases. The wide clinical application of nucleic acid materials dependents on the development of biocompatible gene carriers. There are enormous various compounds widely investigated to be used as non-viral gene carriers including lipids, polymers, carbon materials, and inorganic structures. In this review, we will discuss the recent discoveries on non-viral gene delivery systems. We will also highlight the in vivo gene delivery mediated by non-viral vectors to treat cancer in different tissue and organs including brain, breast, lung, liver, stomach, and prostate. Finally, we will delineate the state-of-the-art and promising perspective of in vivo gene editing using non-viral nano-vectors.

Since the elucidation of the molecular mechanisms of several diseases along with the discovery of nucleic acid structure, the replacement of defective genes with functional versions has been considered as a new therapeutic paradigm called "gene therapy" (1, 2). Gene therapy is carried out by expression constructs in order to increase the production of specific proteins inside the cells. On the other hand, down-regulation of specific genes has shown great potential for the treatment of various diseases (3) . Therefore, the modulation or silencing of such genes using accessed by the ribosome for the production of proteins. On the other hand, mRNA directly enters the cytoplasm and interacts with ribosome for protein production. These unique properties have made mRNA as a potential candidate not only for gene therapy but also for vaccine development particularly for the immunization against widespread viruses including SARS-CoV-2 (36). However, the major concerns regarding the application of mRNA for gene therapy are its unstable nature and the existence of degrading enzymes such as RNases in the extra-and intra-cellular environments (37, 38). To overcome these problems, new developments, including SNIM (stabilized non-immunogenic mRNA), have been introduced in which the modified nucleotides could be incorporated into the mRNA structure to increase its stability and reduce its immunogenicity (4, 39-41).

The aim of gene therapy is not just increasing the expression of certain gene as it was expected in previous decades. There are several pathological conditions related to the genes over-expression.

In such conditions, the gene therapy goal would be silencing the target genes. The knock-down of such genes could be achieved by different nucleic acid materials, including antisense and siRNA. It must be considered that there are some differences between gene therapy and oligonucleotide therapy (42). Oligonucleotide-based medications such as antisense do not need the transcriptional and translational machinery of the cells while the conventional gene therapy is based on the replacement of defected genes by the functional ones as well as the introduction of new gene into the cells including germlines or somatic cells (43). Antisense technology is defined as a powerful tool to down-regulate a specific gene by transferring the antisense strand to the cells with the ability to interact with the sense strand. The base pairing between the sense and antisense strands results in the translational block (44, 45) . On the other hand, RNAi technology employs several enzymes (e.g., dicer) and proteins (e.g., RISC complex) to interfere with the layer on the surface of the carriers reduces the risk of aggregation and increases colloidal stability. The reduction of the interaction between the stealth gene carriers and serum components reduces the recognition of the vehicles by mononuclear phagocyte system (MPS), including macrophages, which in turn leads to enhanced circulation time (58) . In order to direct the carriers into the precise site of action, smart gene carriers have been designed. These carriers could be targeted to the specific receptors by the conjugation of small molecules as well as macromolecules including monoclonal antibodies or aptamers (59) (60) (61) . Once the nano-carriers reach the cells, they may enter endosomal compartment, which degrades the nucleic acid therapeutics and leads to failed transfection. Hence, the promotion of proton sponge effect or the conjugation of membrane fusogenic compounds could be considered as brilliant strategies to overcome the endo/lysosomal barrier (62, 63) . While the siRNA site of action is the cytosolic environment, plasmids must be able to cross the nuclear barrier. It has been shown that the molecules with the molecular mass of 40-70 kDa (10-25 nm) are able to passively diffuse via nuclear pores. However, the exact mechanism of nuclear entry is not completely understood (4, 64) . It is not clear whether the polyplexes goes under vector unpackaging outside the nucleus or the transcriptional machinery of the cell dissociate the nucleic acids from the carrier inside the nucleus. Regardless of the mechanism, it has been demonstrated that cell cycle may have a crucial impact on the cell entry. The cells at the phases of S/G2 have shown the highest transfection efficiency. However, most cells are not in the dividing phase in vivo; therefore the alternative approaches, including the conjugation of nuclear localization signals (NLS), must be employed to increase nucleus entry (65, 66) . The real value of these important findings is dependent on their translation to clinical application. The approval of patisiran (Onpattro ® ) as the first FDA approved siRNA based therapeutic for hereditary transthyretin-mediated (hATTR) J o u r n a l P r e -p r o o f amyloidosis opened up new horizons for the scientists to seek for the efficient delivery systems enabling the nucleic acids to be used as therapeutic agents. Patisiran has been formulated as lipid nanoparticles (NPs) and is used by intravenous infusions while the second approval for siRNAbased therapeutics belongs to givosiran (Givlaari ® ) (67) (68) (69) . Givosiran has been prepared as Nacetylgalactosamine (GalNAc) conjugated siRNA and is administrated subcutaneously. The first polymer-based gene therapy investigation in human was carried by Transferrin-polylysine (adenovirus-enhanced transferrinfection; AVET) carrier in order to transfer the plasmid encoding IL-2 gene for the treatment of melanoma (70) . In the first-ever human study of polyplexes, the ex-vivo gene transfer was performed to deliver the plasmid DNA into the patient cells. PEG conjugated polylysine was used to transfer the pDNA to treat cystic fibrosis as a nasal drug delivery system (16) . In another study to design a vaccine for HIV, mannose conjugated PEI was prepared as the carrier for the plasmid encoding various HIV antigens and used as a dermal formulation in a human clinical trials (71). The intraperitoneal injection of PEG-PEI-Cholesterol to transfer IL-12 plasmid was also used for ovarian cancer treatment (72). The intravenous injection of transferrin-cyclodextrin oligocation complexed with siRNA to silence ribonucleotide reductase M2(RRM2) was applied in various solid tumors (22) . Since various routes of administration have been used to transfer non-viral delivery systems for gene therapy, it seems that the route is highly dependent on the characteristics of the carrier and nucleic acids as well the prepared complex and the final formulation. It seems that there is no restrict limitation for a specific route of administration for non-viral gene delivery carriers at least in the theoretical aspect ( Table 2 ).

J o u r n a l P r e -p r o o f 

Despite advances in chemotherapy, surgery, and radiation therapy, lung cancer is one of the leading causes of cancer-related deaths globally (148, 149) . Even though there is some initial response with present conventional chemotherapy, patients will develop resistance and exhibit poor survival with prolonged usage (150) . Several attempts were made to improve the survival of lung cancer patients using various combination therapies that have demonstrated that no further improvement observed, suggesting the need for specific, less toxic treatment approaches such as genetic alterations. Tumor suppressor genes and oncogenes are the two major genetic factors affecting the progression of the disease (151, 152) . Hence, altering these explicit genes can advance the therapeutic benefit of present therapies. (153) . Numerous gene therapy strategies have been adopted, such as the deletions of oncogenes, immune stimulation, replacement of tumor-suppressor genes and transfer of genes that enhance conventional treatments (154) . Here, There are several other non-viral vectors used for the delivery of various nucleic acid materials for lung cancer (166) (167) (168) (169) (170) (171) (172) (173) . Another most common genetic alteration happen in the lung cancer is associated with the tumor suppressor genes. For example, tumor suppressor gene TUSC2/FUS1

(TUSC2) is inactivated in lung cancer. However, no drug development approach is available for targeting the loss-of-function genetic deviations. Roth JA and his team developed a systemic gene therapy approach by using a TUSC2-expressing plasmid vector packaged in DOTAP:chol nanovesicles. They found that following the tumor treatment with DC-TUSC2, some major changes in the intrinsic pro-apoptotic pathway happened (174, 175) . These nanovesicles were administered intravenously in the patients bearing lung cancer and the results showed an improvement in delivering TUSC2 genes to both human primary and metastatic tumors safely (176) . Among several existing polymeric transporters, PEI was mostly exploited to transfer genes for both in vitro and in vivo transfection. For example, scientists used PEI to develop a pHsensitive in vivo selective gene delivery system to transfer p53DNA at the tumor site. A single administration of p53DNA nanocomplex along with laser radiation, significantly inhibited tumor growth and prolonged median survival (177) . Gold NPs also used to deliver p53DNA to lung cancer cells (178) . Several other studies also demonstrated that the p53-based gene delivery is able to improve the therapeutic outcome for lung cancer (179) (180) (181) (182) . In summary, based on these 

There are several strategies to treat breast cancers based on the severity and the mechanisms involved in the pathogenesis including autophagy and apoptosis (183) . Although there are CXCR4siRNA and an RNA-triple-helix in the hydrogels NPs without synthetic polycationic reagents for the treatment of breast cancer (189) . Amorphous calcium carbonate fusion nanospheres fabricated with CaIP 6 NPs were efficient in carrying genes to the tumor site.

Scientists showed that AKT1 siRNA loaded CaCO3/CaIP 6 nanocomplexes substantially inhibited tumor growth (190) . Similarly, a polypeptide containing LAH4-L1-siMDR1 loaded nanocomplexes displayed significant tumor growth inhibition when used along with PTX. In this study, high MDR1 gene silencing efficacy was observed in the tumor-bearing nude mice (98) .

Enormous efforts are still underway for developing novel and effective gene delivery systems based on biocompatible nanomaterials to transfer the target genes to the tumor site (167, 191, 192) . For example, researchers have developed an elastin-like recombinant (ELR) and specific MUC1 aptamers for intracellular delivery of the MUC1 gene to breast tumors (193) . More recently, the same group developed a double protection tumor-specific nanomaterial device for gene therapy in breast cancer (86) . The functionalized peptides/ligands can also improve the delivery of nucleic acid-complexed NPs to tumors (95, 194, 195) . (196) . Cell-penetrating peptide (CPP)-containing and EGFR-siRNA loaded nanobubbles showed synergism with ultrasound irradiation mediated EGFR-siRNA delivery to TNBC (197) . Zhou et al also developed CD105-conjugated targeted cationic microbubbles for antiangiogenesis gene therapy for breast cancer (198) . Similarly, endostatin loaded and CD105 antibody conjugated immunoliposomes were prepared for antiangiogenic and imaging therapy (199) . Gu nanocomplexes. This nanocomplex could inhibited the tumor growth synergistically and prolonged the survival of drug-resistant breast tumors mice (202) . In another similar study, the investigators proved that the co-delivery of p53 DNA and AVPI peptide enabled a complete arrest of tumor growth when used in combination with a reduced dose of Dox. In their study, they modified AVPI peptide not only to enable it to penetrate to tumor cells but also acts as a gene delivery vehicle by forming a nano complex with cationic R8 moiety (203) . There are J o u r n a l P r e -p r o o f several studies demonstrating that the p53 mediated gene therapy for breast cancer treatment is an efficient approach in cancer gene therapy (204, 205) . Overall, the combination of chemotherapy along with gene therapy may enhance the therapeutic effects against breast cancer.

There are other categorization methods for brain tumors including primary and secondary tumors. Primary tumors originate from meninges, glands, nerve and other brain cells, while secondary tumors originate from other parts of the body and spread to the brain (206) . The most common brain cancers are glioma, neuroblastoma, meningioma, vestibular schwannoma and pituitary adenoma. The brain tumors can be primary diagnosed using MRI, CT scan, angiography, skull X-ray and biopsy. Despite enormous advances in the field of pharmaceutics and radiotherapy, the brain cancers cannot be completely cured.

Polymer-based carriers are accounted as one of the most effective carriers in drug delivery (207) (208) (209) (210, 211) . Moreover, gene delivery is accounted as a hopeful strategy for brain cancer treatment. One of the most important obstacles in brain drug delivery is the blood-brain barrier (BBB). Therefore, there are many efforts to overcome this barrier including functionalization and modification of non-viral gene delivery vectors (212, 213) . The modification leads to the transcytosis and endocytosis of vectors through cell-penetrating peptides (CPP) mediated transmembrane transport, adsorptive-mediated endocytosis and receptor-mediated endocytosis (214) . There are some receptors on the surface of it seems that R7L10 is safer than PEI and conjugation with Epo enhances its gene and drug delivery efficacy in hypoxia condition (107) .

Dendrimers have been considered as effective drug delivery carriers and polyamidoamine (PAMAM) is one the most well-known dendrimers in drug delivery. It seems that primary and tertiary amines in dendrimer play a critical role in DNA condensation and release (226) .

However, there are controversial reports on the safety of dendrimers owing to their positive surface charge, especially for G2-G4 dendrimers (227, 228) . It has been shown that PEGylated lactoferrin-dendrimer-DNA has shown significantly less toxicity and higher transfection efficacy than non-PEGylated ones. Interestingly, they showed that brain uptake and transfection efficacy of the lactoferrin conjugated complexes were significantly higher than the transferrin substituted enhances cell viability, rat survival and VEGF marker while decreases apoptosis as compared to the polyplex-TRAIL, polyplex-HSV-TK and PBS in glioma-bearing SD rats (99) . However, the complex containing SV-TK with erythropoietin and nestin intron 2 (NI2) showed that its complexation with reducible poly oligo D-arginine has significantly less cytotoxicity than PEI even at hypoxic condition. Furthermore, the polyplex induced significantly higher apoptosis and tumor size decrease in an intracranial glioblastoma rat model (106) . Overall, the targeting strategies might be considered as a prerequisite for non-viral vectors used for brain gene therapy.

J o u r n a l P r e -p r o o f

gastrointestinal cancers including colorectal and gastric cancers. These nano carriers have been employed as delivery vehicles for RNA silencing of oncogenes, DNA delivery of tumor suppressors, apoptosis inducers, suicide genes or immune-stimulatory molecules.

Colorectal cancer is the third most deadly diagnosed cancer in the world due to its metastasis tumor-bearing nude mice (266) .

In addition to various materials used for the delivery of nucleic acids for colorectal carcinoma, electrotransfection is a promising route for facilitated delivery of genes into the target cells. 

Gastric cancer is the second most malignant cancer worldwide with the poor five-year survival of 30% (267) . A range of nanoparticulate systems have been investigated for efficient and safe delivery of genes to gastric cancer models. For example, calcium phosphate NPs (CPNPs) were used to deliver a novel fusion suicide gene, yCDglyTK, which is regulated by a cancer-specific CEA promoter and a CMV enhancer (CV) (268) (269) (270) . It was observed that CPNPs specifically 

Hepatocellular carcinoma (HCC) is another deadly cancer worldwide due to the late diagnosis and the impaired and insufficient treatments. Therefore, it is necessary to develop the carriers with enhanced targeted specificity, improved efficiency and safety (276, 277) . its promoter is associated with the risk of‫‬ several cancers including HCC. It can also be a predictive factor for poor HCC prognosis. In vivo efforts to re-express RASSF1A has shown the arrest of HCC growth as well as the improved sensitivity of HCC cells to mitomycin (282) . plasmid DNA was used as a reporter gene. The average hydrodynamic size and zeta potential of the carrier system at C/P ratio of 25 were 157 ± 3 nm and +18 ± 0.3 mV, respectively. The nanovehicle was intratumorally injected to subcutaneous Huh-7 xenografts in athymic nude mice. It was suggested that biodegradable 536 NPs would also be appropriate for systemic or transarterial delivery due to its small size which preferentially localized in tumor through EPR effect (126) .

In another effort for HCC gene therapy, a multifunctional NP targeted for HCC was designed to deliver TRAIL gene in mice (251) . These self-assembled lipid-bilayer structures (LCPP NPs) are composed of the calcium phosphate (CaP) and protamine core, which act as a pH stimuliresponsive and TRAIL nuclear localization agent, respectively. Moreover, The Ca ions released from CaP reverse the TRAIL resistance. HCC-targeting peptide (SP94) was also used for for hepatocellular carcinoma treatment. The results showed that the treatment of the cells with such system combined with ultrasound irradiation increased the miR-122 expression level by 30fold in human HCC xenografts (130) . Hence, these methods have shown potential for further studies to develop safe and efficient gene therapy approaches.

Prostate cancer is the fourth most common cancer and the second most extensive cancer in males leading to the mortality of around 300,000 individuals per year. Almost 200,000 new patients have been diagnosed per annum. The late diagnosis of prostate cancer is the primary cause of death (290, 291) . Based on the stage and severity of the tumor, different treatments can be suggested to the patient including prostatectomy, radiotherapy, hormone therapy, chemotherapy, gene therapy, and a combination of them. The most recent procedure is gene therapy that mainly initiated via transferring a new gene to achieve destruction or fixation of cancerous cells (292) (293) (294) . Transferrin and lactoferrin are two iron-binding proteins that widely used as targeting ligands for prostate cancers (295, 296) . Another promising approach for prostate targeting is using the integrins that can be attached to the extracellular matrix of prostate cancer J o u r n a l P r e -p r o o f microenvironment. Integrin receptors are supposed to be over-expressed on prostate cancer cells (297, 298) . Prostate-specific membrane antigen (PSMA), integrins, and prostate stem cell antigen (PSCA) are the glycoprotein which could be targeted by various ligands (299, 300) . 

The most common treatment of cancers is chemotherapy while having various challenges and side effects including the lack of selectivity to the cancer cells and toxicity to the healthy cells (305) . Different approaches such as gene therapy and combination therapy have been suggested to circumvent these limitations (111, 285, 306) . Combination therapy may decrease the toxicity of each agent by reducing the individual drug-related dose. In this field, co-delivery of drug and gene-based NPs have attracted more attention (244, 245) . co-delivery of doxorubicin and siRNA against P-glycoprotein. It has been observed that the emerged synergistic effect is even more efficient than co-treatment of chemotherapeutics and siRNA (116, 193, 244, 285) . Only in these systems, Cas9 protein is an essential compartment for DNA interference.

Generally, this system contains a nuclease protein (Cas9) and a guide RNA (gRNA) (213) .

Since, the gRNA could be replaced by sgRNA (synthetic chimeric single guide RNA), the Cas9 protein could be directed to the target site using sgRNA which consequently leads to the induction of double-stranded DNA breaks (DSBs). Finally, the major pathways of repair mechanism in the cells are responsible for inducing the alterations. This simple, robust, userfriendly, specific, and efficient system has enabled researchers to create models for various diseases as well the novel therapeutic approaches (314) (315) (316) (317) .

Generally, there are three different approaches for CRISPR/Cas 9 delivery (318) . The ultimate goal is to transfer the whole system into the cells. However, the ribonucleoprotein complex could Since the difficulties for efficient delivery of Cas9 protein reduces the transfection efficiency of sgRNA and Cas9 protein, the alternative strategy is to use the Cas9 mRNA with sgRNA. For efficient delivery of Cas9 protein and sgRNA, the delivery platform must be able to transfer a large positively charged protein (Cas9) and a negatively charged nucleic acid (sgRNA) together.

Designing such delivery systems is not simple. The second approach includes the delivery of two mRNA molecules with similar biophysical properties that facilitates the design of delivery systems. Besides, the introduction of Cas9 mRNA into cells does not need to be entered into the cell nucleus for subsequent transcription. Therefore, the main advantage for this approach is the quick onset of action. The transient expression of Cas9 mRNA in the cytosol along with the quick onset of Cas9 action make this approach an attractive way for the researchers to reduce the off-target effects associated with the long time presence of Cas9 protein inside the cells.

However, the low stability of mRNA is the major hampering factor for this delivery method.

Various non-viral delivery strategies have been employed to transfer Cas9 mRNA with the sgRNA together including zwitterionic aminolipid NPs (332-335) and branched-tail lipid NPs.

Since there are several problems for efficient delivery of Cas9 protein, Cas 9 mRNA and sgRNA, the third method have been introduced which includes the design of a plasmid encoding Cas 9 and sgRNA inside the cells. The stability of plasmid-based CRISPR/Cas9 systems is really higher than protein or mRNA making these systems more attractive for in vivo applications.

However, there are several major obstacles reducing its clinical applications. This system could be able to cross the nuclear membrane and access the transcriptional machinery of the cells.

Since the transcription of the plasmids and the production of Cas9 protein and sgRNA need more time rather than the direct introduction of these macromolecules into the cells, the delay in the onset of therapeutic action is expected. In addition, the off-target effects associated with the long-term production of Cas9 protein is more probable rather than the previous methods. Also, the risk of the integration of plasmid into the genomic materials may reduce their potential for 

In recent decades, various oligonucleotide-based therapeutics have been introduced for human clinical applications. This novel category of therapeutic materials includes antisense oligonucleotides and aptamers as well as siRNA-based medications. The clinical applications of these new drugs are the result of breakthrough discoveries in molecular biology. However, the translation of these achievements to the clinical applications is substantially dependent on the development of efficient and safe delivery systems. An optimized delivery system for nucleic acids should be able to form a stable structure outside the cells and release the payloads at the specific site of action. In addition, the toxicity of the delivery vehicle must be tolerable by the human cells. The biophysical properties and the pharmacokinetic characteristics of the vehicles are the other significant points which determine the potential of delivery system for human applications. In order to improve these properties, stealth technology using various materials such as PEG and targeting strategies have been introduced. Using these approaches, the biophysical characteristics of the carriers could be modified and their pharmacokinetic properties might be improved. Generally, polymer and dendrimer-based delivery systems have shown higher transfection efficiency (5, 25, 53) . However, their toxicity is the major concern for the further developments towards the clinical applications. For these carrier systems, the main J o u r n a l P r e -p r o o f modification strategy is focused on the reduction of cytotoxicity through the modulation of cationic charge or designs the biodegradable polycationic compounds. In addition, these materials suffer from the low targetability for the specific cells or tissues (346). Therefore, the addition of targeting moieties on these materials could be considered as an effective way to improve their properties. These materials are appropriate delivery systems for the formation of complexes based on the electrostatic interaction between the nucleic acid and carrier. On the other hand, lipid-based carriers have demonstrated higher biocompatibility rather than the polymeric delivery systems (347, 348). These delivery systems have shown great potential for clinical applications due to their low toxicity. However, the transfection efficiency of such materials is generally lower than the polymeric compounds. Therefore, the major approaches to improve the properties of these vehicles are focused on the augmentation of their transfection efficiency. Similar to the polymeric delivery systems, lipid-based materials need the targeting moieties for efficient transfer of nucleic acid to the target cells or organs. Although the toxicity of lipid-based delivery systems is lower than the polycationic polymers or dendrimers, they may induce inflammatory responses following systemic administration. The translation of these materials for commercial application needs a scalable production process which leads to the commercial products with highest batch-to-batch uniformity. The most recent clinical trial on the application of mRNA as a potential vaccine for SARS-CoV-2 has been conducted by LNPs which shows the importance of this category of delivery system for human application (36 

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Nanoghosts as a Novel Natural Nonviral Gene Delivery Platform Safely Targeting Multiple Cancers

Study on the prostate cancer-targeting mechanism of aptamer-modified nanoparticles and their potential anticancer effect in vivo

Highly efficient cationic hydroxyethylated cholesterol-based nanoparticle-mediated gene transfer in vivo and in vitro in prostate carcinoma PC-3 cells

Cancer statistics

Global Epidemiology of Lung Cancer

Emerging insights of tumor heterogeneity and drug resistance mechanisms in lung cancer targeted therapy

Hallmarks of Cancer: The Next Generation

Revisiting the hallmarks of cancer

State-of-the-art human gene therapy: part II. Gene therapy strategies and clinical applications

Gene Therapy Leaves a Vicious Cycle

Non Viral Vectors in Gene Therapy-An Overview

Viral and Non-viral Vectors in Gene Therapy: Technology Development and Clinical Trials

Production and clinical development of nanoparticles for gene delivery

The Development of Functional Non-Viral Vectors for Gene Delivery

siRNA Conjugated Nanoparticles-A Next Generation Strategy to Treat Lung Cancer

Aptamer-targeted delivery of Bcl-xL shRNA using alkyl modified PAMAM dendrimers into lung cancer cells

PolyMetformin combines carrier and anticancer activities for in vivo siRNA delivery

Multi-functional self-assembled nanoparticles for pVEGF-shRNA loading and anti-tumor targeted therapy

Folate-conjugated polyspermine for lung cancer-targeted gene therapy

Exosome-mediated microRNA-497 delivery for anticancer therapy in a microfluidic 3D lung cancer model

MDM2 knockdown mediated by a triazinemodified dendrimer in the treatment of non-small cell lung cancer

AT2R Gene Delivered by Condensed Polylysine Complexes Attenuates Lewis Lung Carcinoma after Intravenous Injection or Intratracheal Spray

Elastin-like recombinamers with acquired functionalities for gene-delivery applications

Cationic lipid guided shorthairpin RNA interference of annexin A2 attenuates tumor growth and metastasis in a mouse lung cancer stem cell model

Preparation and evaluation of chitosan-DNA-FAP-B nanoparticles as a novel non-viral vector for gene delivery to the lung epithelial cells

Visualization and Expression of Genes with Biomimetically Mineralized Zeolitic Imidazolate Framework-8 (ZIF-8)

Therapeutic Effect of pHLIP-mediated CEACAM6 Gene Silencing in Lung Adenocarcinoma

Tumor Suppressor FUS1 Signaling Pathway

Synergistic Tumor Suppression by Coexpression of FUS1 and p53 Is Associated with Down-regulation of Murine Double Minute-2 and Activation of the Apoptotic Protease-Activating Factor 1-Dependent Apoptotic Pathway in Human Non-Small Cell Lung Cancer Cells

Phase I Clinical Trial of Systemically Administered TUSC2(FUS1)-Nanoparticles Mediating Functional Gene Transfer in Humans

Highly specific in vivo gene delivery for p53-mediated apoptosis and genetic photodynamic therapies of tumour

<p>Fabrication Of Gold Nanoparticles In Absence Of Surfactant As In Vitro Carrier Of Plasmid DNA</p>

Poly( -amino ester) Nanoparticle Delivery of TP53 Has Activity against Small Cell Lung Cancer In Vitro and In Vivo

Current Status of Gene Therapy for Lung Cancer and Head and Neck Cancer

Histone H2A-peptide-hybrided upconversion mesoporous silica nanoparticles for bortezomib/p53 delivery and apoptosis induction

Autophagic, apoptotic, and necrotic cancer cell fates triggered by acidic pH microenvironment

Toxicity concerns of nanocarriers. Nanotechnology-Based Approaches for Targeting and Delivery of Drugs and Genes

Hydrogel Doped with Nanoparticles for Local Sustained Release of siRNA in Breast Cancer

Direct cytosolic siRNA delivery by reconstituted high density lipoprotein for target-specific therapy of tumor angiogenesis

Codelivery of an Optimal Drug/siRNA Combination Using Mesoporous Silica Nanoparticles To Overcome Drug Resistance in Breast Cancer in Vitro and in Vivo

TPGS functionalized mesoporous silica nanoparticles for anticancer drug delivery to overcome multidrug resistance

A self-assembled RNA-triple helix hydrogel drug delivery system targeting triple-negative breast cancer

CaCO3/CaIP6 composite nanoparticles effectively deliver AKT1 small interfering RNA to inhibit human breast cancer growth

Elastin-Like Recombinamers As Smart Drug Delivery Systems

Biocompatibility and immunogenicity of elastin-like recombinamer biomaterials in mouse models

Biocompatible ELR-Based Polyplexes Coated with MUC1 Specific Aptamers and Targeted for Breast Cancer Gene Therapy

Engineering Nanoparticles for Targeted Delivery of Nucleic Acid Therapeutics in Tumor

Cell Penetrating Peptides, Novel Vectors for Gene Therapy

Synthesis of a novel PEGDGA-coated hPAMAM complex as an efficient and biocompatible gene delivery vector: an in vitro and in vivo study

Novel cell-penetrating peptide-loaded nanobubbles synergized with ultrasound irradiation enhance EGFR siRNA delivery for triple negative Breast cancer therapy

Targeted Antiangiogenesis Gene Therapy Using Targeted Cationic Microbubbles Conjugated with CD105 Antibody Compared with Untargeted Cationic and Neutral Microbubbles

Efficient targeted tumor imaging and secreted endostatin gene delivery by anti-CD105 immunoliposomes

Reversal of P-glycoprotein-mediated multidrug resistance by CD44 antibody-targeted nanocomplexes for short hairpin RNA-encoding plasmid DNA delivery

Polycation-functionalized nanoporous silicon particles for gene silencing on breast cancer cells

A cationic prodrug/therapeutic gene nanocomplex for the synergistic treatment of tumors

DNA Nanocomplex as Adjuvant Therapy for Drug-Resistant Breast Cancer

Comparison of two silica based nonviral gene therapy vectors for breast carcinoma: evaluation of the p53 delivery system in Balb/c mice

Novel luminescent silica nanoparticles (LSN): p53 gene delivery system in breast cancer in vitro and in vivo

Systematic Study of NaF Nanoparticles in Micelles loaded on Polylactic Acid Nanoscaffolds: In Vitro Efficient Delivery

A Systematic Study of Cu Nanospheres Embedded in Nonionic Surfactant-Based Vesicle: Photocatalytic Efficiency and In Vivo Imaging Study

Analysis of nanoparticle delivery to tumours

Progress in Microneedle-Mediated Protein Delivery

Multifunctional Polymeric Nanoplatforms for Brain Diseases Diagnosis

ZEB1 and ZEB2 gene editing mediated by CRISPR/Cas9 in A549 cell line

Non-viral gene delivery and therapeutics targeting to brain

Quantitative evaluation of monocyte transmigration into the brain following chemical opening of the blood-brain barrier in mice

Non-viral vectors based on cationic niosomes as efficient gene delivery vehicles to central nervous system cells into the brain

OX26/CTX-conjugated PEGylated liposome as a dual-targeting gene delivery system for brain glioma

Nanogels for oligonucleotide delivery to the brain

A powerful nonviral vector for in vivo gene transfer into the adult mammalian brain: polyethylenimine. Human gene therapy

Dual-targeting and microenvironmentresponsive micelles as a gene delivery system to improve the sensitivity of glioma to radiotherapy

Boosting RNAi therapy for orthotopic glioblastoma with nontoxic brain-targeting chimaeric polymersomes

Growth inhibition, cytokinesis failure and apoptosis of multidrug-resistant leukemia cells after treatment with P-glycoprotein inhibitory agents

Stability, intracellular delivery, and release of siRNA from chitosan nanoparticles using different cross-linkers

Receptor-mediated gene delivery using PAMAM dendrimers conjugated with peptides recognized by mesenchymal stem cells

Protective effect of PEGylation against poly

Glioblastoma U-87MG tumour cells suppressed by ZnO folic acid-conjugated nanoparticles: an in vitro study. Artificial cells, nanomedicine, and biotechnology

Preparation, characterization, and evaluation of the anticancer activity of artemether-loaded nanoniosomes against breast cancer

Improved drug delivery and therapeutic efficacy of PEgylated liposomal doxorubicin by targeting anti-HER2 peptide in murine breast tumor model

Targeted lung cancer therapy: preparation and optimization of transferrin-decorated nanostructured lipid carriers as novel nanomedicine for codelivery of anticancer drugs and DNA

Biocompatible ELR-based polyplexes

Biodegradable poly (ɛ-caprolactone)-poly (ethylene glycol) copolymers as drug delivery system

PCL/PEG copolymeric nanoparticles: potential nanoplatforms for anticancer agent delivery

Cationic micelle-based siRNA delivery for efficient colon cancer gene therapy

Treating colon cancer with a suicide gene delivered by self-assembled cationic MPEG-PCL micelles

Novel polymer micelle mediated codelivery of doxorubicin and P-glycoprotein siRNA for reversal of multidrug resistance and synergistic tumor therapy

A multifunctional nanocarrier for TRAI -b herapy against hepatocellular carcinoma with desmoplasia in mice

Survivin-T34A: molecular mechanism and therapeutic potential

Regulation of apoptosis at cell division by p34cdc2 phosphorylation of survivin

Nanotechnology-based siRNA delivery strategies for metastatic colorectal cancer therapy

Nanoparticle-based delivery of siDCAMKL-1 increases microRNA-144 and inhibits colorectal cancer tumor growth via a Notch-1 dependent mechanism

Selective blockade of DCAMKL-1 results in tumor growth arrest by a Let-7a MicroRNA-dependent mechanism

Cell state plasticity, stem cells, EMT, and the generation of intra-tumoral heterogeneity

Synthetic anticancer gene medicine exploits intrinsic antitumor activity of cationic vector to cure established tumors

Nan Fang Yi Ke Da Xue Xue Bao

Novel polyethyleneimine-R8-heparin nanogel for high-efficiency gene delivery in vitro and in vivo

Efficient inhibition of C-26 colon carcinoma by VSVMP gene delivered by biodegradable cationic nanogel derived from polyethyleneimine

Synergistic and low adverse effect cancer immunotherapy by immunogenic chemotherapy and locally expressed PD-L1 trap

Liposomal nanostructures for drug delivery in gastrointestinal cancers

Liposome-encapsulated plasmid DNA of telomerase-specific oncolytic adenovirus with stealth effect on the immune system

Cationic liposome coupled endostatin gene for treatment of peritoneal colon cancer

Nanoparticles to Deal with Gastric Cancer

Suicide gene delivery by calcium phosphate nanoparticles: a novel method of targeted therapy for gastric cancer

Calcium phosphate nanoparticles as a novel nonviral vector for efficient transfection of DNA in cancer gene therapy

Tissue specific expression of suicide genes delivered by nanoparticles inhibits gastric carcinoma growth

Regression of gastric cancer by systemic injection of RNA nanoparticles carrying both ligand and siRNA

Photothermal and gene therapy combined with immunotherapy to gastric cancer by the gold nanoshell-based system

Novel polyethylenimine-derived nanoparticles for in vivo gene delivery

Development of an MRI-visible nonviral vector for siRNA delivery targeting gastric cancer

Characterization of polyethylene glycolgrafted polyethylenimine and superparamagnetic iron oxide nanoparticles (PEG-g-PEI-SPION) as an MRIvisible vector for siRNA delivery in gastric cancer in vitro and in vivo

Current status of nanomaterial-based treatment for hepatocellular carcinoma

Advances in delivery vectors for gene therapy in liver cancer

Gene therapy approaches against cancer using in vivo and ex vivo gene transfer of interleukin-12

Asialoglycoprotein receptor-magnetic dual targeting nanoparticles for delivery of RASSF1A to hepatocellular carcinoma

RASSF1A expression inhibits the growth of hepatocellular carcinoma from Qidong County

A nanoparticle-based model delivery system to guide the rational design of gene delivery to the liver. 2. In vitro and in vivo uptake results

Gold nanoparticles delivered miR-375 for treatment of hepatocellular carcinoma

Advances in the application of nanotechnology in the diagnosis and treatment of gastrointestinal tumors

PTEN and TRAIL genes loaded zein nanoparticles as potential therapy for hepatocellular carcinoma

m v αfetoprotein promoter-mediated tBid delivered by folate-PEI600-cyclodextrin nanopolymer vector in hepatocellular carcinoma

YAP inhibition restores hepatocyte differentiation in advanced HCC, leading to tumor regression

Highly efficient and tumorselective nanoparticles for dual-targeted immunogene therapy against cancer

Prostate cancer gene therapy clinical trials

Prostate cancer: diagnosis and clinical management

History of gene therapy

Non-viral gene delivery methods

Cancer nanotechnology: the impact of passive and active targeting in the era of modern cancer biology

Regression of prostate tumors after intravenous administration of lactoferrin-bearing polypropylenimine dendriplexes encoding TNF-α TRAI k -12

Therapeutic efficacy of intravenously administered transferrin-conjugated dendriplexes on prostate carcinomas

Integrins and prostate cancer metastases

Tumor targeting via integrin ligands

Novel Strategies for Targeting Prostate Cancer

Targeted nonviral gene therapy in prostate cancer

Preparation of nanobubbles carrying androgen receptor siRNA and their inhibitory effects on androgen-independent prostate cancer when combined with ultrasonic irradiation

Interleukin-27 gene delivery for modifying malignant interactions between prostate tumor and bone

DNA/Lipid complex incorporated with fibronectin to cell adhesion enhances transfection efficiency in prostate cancer cells and xenografts

Autophagy modulators: mechanistic aspects and drug delivery systems

Which one performs better for targeted lung cancer combination therapy: pre-or postbombesin-decorated nanostructured lipid carriers? Drug delivery

Near-infrared/pH dual-responsive nanocomplexes for targeted imaging and chemo/gene/photothermal tri-therapies of non-small cell lung cancer

Nanotechnological Strategies for Osteoarthritis Diagnosis, Monitoring, Clinical Management, and Regenerative Medicine: Recent Advances and Future Opportunities

Memory of viral infections by CRISPR-Cas adaptive immune systems: acquisition of new information

RNA-guided genetic silencing systems in bacteria and archaea

EMT signaling: potential contribution of CRISPR/Cas gene editing

Multiplexed CRISPR technologies for gene editing and transcriptional regulation

Efficient genome editing in zebrafish using a CRISPR-Cas system

Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials

Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells

Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease

Harnessing nanoparticles for the efficient delivery of the CRISPR/Cas9 system

Direct cytosolic delivery of CRISPR/Cas9-ribonucleoprotein for efficient gene editing

Cytosolic delivery of CRISPR/Cas9 ribonucleoproteins for genome editing using chitosan-coated red fluorescent protein

Gene disruption by cellpenetrating peptide-mediated delivery of Cas9 protein and guide RNA

A pH-responsive silica-metalorganic framework hybrid nanoparticle for the delivery of hydrophilic drugs, nucleic acids, and CRISPR-Cas9 genome-editing machineries

A biodegradable nanocapsule delivers a Cas9 ribonucleoprotein complex for in vivo genome editing

The authors would like to thank Dr. Horacio Cabral (Department of Materials Engineering, The University of Tokyo) for his constructive comments.