Doxorubicin Improves Cancer Cell Targeting by Filamentous Phage Gene Delivery Vectors

Merging targeted systemic gene delivery and systemic chemotherapy against cancer, chemovirotherapy, has the potential to improve chemotherapy and gene therapy treatments and overcome cancer resistance. We introduced a bacteriophage (phage) vector, named human adeno-associated virus (AAV)/phage or AAVP, for the systemic targeting of therapeutic genes to cancer. The vector was designed as a hybrid between a recombinant adeno-associated virus genome (rAAV) and a filamentous phage capsid. To achieve tumor targeting, we displayed on the phage capsid the double-cyclic CDCRGDCFC (RGD4C) ligand that binds the alpha-V/beta-3 (αvβ3) integrin receptor. Here, we investigated a combination of doxorubicin chemotherapeutic drug and targeted gene delivery by the RGD4C/AAVP vector. Firstly, we showed that doxorubicin boosts transgene expression from the RGD4C/AAVP in two-dimensional (2D) cell cultures and three-dimensional (3D) tumor spheres established from human and murine cancer cells, while preserving selective gene delivery by RGD4C/AAVP. Next, we confirmed that doxorubicin does not increase vector attachment to cancer cells nor vector cell entry. In contrast, doxorubicin may alter the intracellular trafficking of the vector by facilitating nuclear accumulation of the RGD4C/AAVP genome through destabilization of the nuclear membrane. Finally, a combination of doxorubicin and RGD4C/AAVP-targeted suicide gene therapy exerts a synergistic effect to destroy human and murine tumor cells in 2D and 3D tumor sphere settings.

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An artificial cell system for biocompatible gene delivery in cancer therapy

Nanoscale ◽

10.1039/c9nr09131a ◽

2020 ◽

Vol 12 (18) ◽

pp. 10189-10195 ◽

Cited By ~ 7

Author(s):

Xin Zhao ◽

Dongyang Tang ◽

Ying Wu ◽

Shaoqing Chen ◽

Cheng Wang

Keyword(s):

Gene Therapy ◽

Cancer Therapy ◽

Gene Delivery ◽

Cancer Cells ◽

Cell System ◽

Artificial Cell

The artifical cell system for the gene therapy of cancer might be a promising approach for the reversal of neoplastic progress of cancer cells.

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Adeno-Associated Viral Vectors for Gene Transfer and Gene Therapy

Biological Chemistry ◽

10.1515/bc.1999.078 ◽

1999 ◽

Vol 380 (6) ◽

Cited By ~ 63

Author(s):

H. Büeler

Keyword(s):

Gene Expression ◽

Gene Therapy ◽

Gene Transfer ◽

Gene Delivery ◽

Viral Vectors ◽

Delivery Systems ◽

Adeno Associated Virus ◽

Viral Genes ◽

Virus Recombinant

AbstractAdeno-associated virus (AAV) is a defective, non-pathogenic human parvovirus that depends for growth on coinfection with a helper adenovirus or herpes virus. Recombinant adeno-associated viruses (rAAVs) have attracted considerable interest as vectors for gene therapy. In contrast to other gene delivery systems, rAAVs lack all viral genes and show long-term gene expression

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Identification and Validation of Small Molecules That Enhance Recombinant Adeno-associated Virus Transduction following High-Throughput Screens

Journal of Virology ◽

10.1128/jvi.02953-15 ◽

2016 ◽

Vol 90 (16) ◽

pp. 7019-7031 ◽

Cited By ~ 22

Author(s):

Sarah C. Nicolson ◽

Chengwen Li ◽

Matthew L. Hirsch ◽

Vincent Setola ◽

R. Jude Samulski

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

High Throughput ◽

Adeno Associated Virus ◽

Virus Gene ◽

Diploid Cells ◽

High Throughput Screens ◽

Group 1

ABSTRACTWhile the recent success of adeno-associated virus (AAV)-mediated gene therapy in clinical trials is promising, challenges still face the widespread applicability of recombinant AAV(rAAV). A major goal is to enhance the transduction efficiency of vectors in order to achieve therapeutic levels of gene expression at a vector dose that is below the immunological response threshold. In an attempt to identify novel compounds that enhance rAAV transduction, we performed two high-throughput screens comprising 2,396 compounds. We identified 13 compounds that were capable of enhancing transduction, of which 12 demonstrated vector-specific effects and 1 could also enhance vector-independent transgene expression. Many of these compounds had similar properties and could be categorized into five groups: epipodophyllotoxins (group 1), inducers of DNA damage (group 2), effectors of epigenetic modification (group 3), anthracyclines (group 4), and proteasome inhibitors (group 5). We optimized dosing for the identified compounds in several immortalized human cell lines as well as normal diploid cells. We found that the group 1 epipodophyllotoxins (teniposide and etoposide) consistently produced the greatest transduction enhancement. We also explored transduction enhancement among single-stranded, self-complementary, and fragment vectors and found that the compounds could impact fragmented rAAV2 transduction to an even greater extent than single-stranded vectors.In vivoanalysis of rAAV2 and all of the clinically relevant compounds revealed that, consistent with ourin vitroresults, teniposide exhibited the greatest level of transduction enhancement. Finally, we explored the capability of teniposide to enhance transduction of fragment vectorsin vivousing an AAV8 capsid that is known to exhibit robust liver tropism. Consistent with ourin vitroresults, teniposide coadministration greatly enhanced fragmented rAAV8 transduction at 48 h and 8 days. This study provides a foundation based on the rAAV small-molecule screen methodology, which is ideally used for more-diverse libraries of compounds that can be tested for potentiating rAAV transduction.IMPORTANCEThis study seeks to enhance the capability of adeno-associated viral vectors for therapeutic gene delivery applicable to the treatment of diverse diseases. To do this, a comprehensive panel of FDA-approved drugs were tested in human cells and in animal models to determine if they increased adeno-associated virus gene delivery. The results demonstrate that particular groups of drugs enhance adeno-associated virus gene delivery by unknown mechanisms. In particular, the enhancement of gene delivery was approximately 50 to 100 times better with than without teniposide, a compound that is also used as chemotherapy for cancer. Collectively, these results highlight the potential for FDA-approved drug enhancement of adeno-associated virus gene therapy, which could result in safe and effective treatments for diverse acquired or genetic diseases.

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Enhancement of Muscle Gene Delivery with Pseudotyped Adeno-Associated Virus Type 5 Correlates with Myoblast Differentiation

Journal of Virology ◽

10.1128/jvi.75.16.7662-7671.2001 ◽

2001 ◽

Vol 75 (16) ◽

pp. 7662-7671 ◽

Cited By ~ 71

Author(s):

Dongsheng Duan ◽

Ziying Yan ◽

Yongping Yue ◽

Wei Ding ◽

John F. Engelhardt

Keyword(s):

Gene Therapy ◽

Clinical Trials ◽

Gene Delivery ◽

Muscle Cells ◽

Therapeutic Benefit ◽

Transduction Efficiency ◽

Great Promise ◽

Adeno Associated Virus ◽

Muscle Gene ◽

Duchenne's Muscular Dystrophy

ABSTRACT Adeno-associated virus (AAV)-based muscle gene therapy has achieved tremendous success in numerous animal models of human diseases. Recent clinical trials with this vector have also demonstrated great promise. However, to achieve therapeutic benefit in patients, large inocula of virus will likely be necessary to establish the required level of transgene expression. For these reasons, efforts aimed at increasing the efficacy of AAV-mediated gene delivery to muscle have the potential for improving the safety and therapeutic benefit in clinical trials. In the present study, we compared the efficiency of gene delivery to mouse muscle cells for recombinant AAV type 2 (rAAV-2) and rAAV-2cap5 (AAV-2 genomes pseudo-packaged into AAV-5 capsids). Despite similar levels of transduction by these two vectors in undifferentiated myoblasts, pseudotyped rAAV-2cap5 demonstrated dramatically enhanced transduction in differentiated myocytes in vitro (>500-fold) and in skeletal muscle in vivo (>200-fold) compared to rAAV-2. Serotype-specific differences in transduction efficiency did not directly correlate with viral binding to muscle cells but rather appeared to involve endocytic or intracellular barriers to infection. Furthermore, application of this pseudotyped virus in a mouse model of Duchenne's muscular dystrophy also demonstrated significantly improved transduction efficiency. These findings should have a significant impact on improving rAAV-mediated gene therapy in muscle.

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High-Effective Gene Delivery Based on Cyclodextrins Multivalent Assembly in Target Cancer Cells

Journal of Materials Chemistry B ◽

10.1039/d1tb02585f ◽

2022 ◽

Author(s):

Yao-Hua Liu ◽

Yu Liu

Keyword(s):

Gene Therapy ◽

Controlled Release ◽

Gene Delivery ◽

Nucleic Acids ◽

Cancer Cells ◽

Chemical Biology ◽

Supramolecular Assembly ◽

Target Cancer

Nucleic acids condensation and controlled release remain significant challenges of gene therapy in chemical biology and nanotechnology fields. In this work, we have reported a polysaccharide supramolecular assembly constructed by...

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Adeno-associated virus-mediated antiapoptotic gene delivery: in vivo gene therapy for neurological disorders

Methods ◽

10.1016/s1046-2023(02)00229-3 ◽

2002 ◽

Vol 28 (2) ◽

pp. 248-252 ◽

Cited By ~ 12

Author(s):

Hideki Mochizuki ◽

Masauki Miura ◽

Takashi Shimada ◽

Yoshikuni Mizuno

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Neurological Disorders ◽

Adeno Associated Virus ◽

Antiapoptotic Gene

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Gene delivery to the hypoglossal motor system: preclinical studies and translational potential

Gene Therapy ◽

10.1038/s41434-021-00225-1 ◽

2021 ◽

Author(s):

Brendan M. Doyle ◽

Michele L. Singer ◽

Thomaz Fleury-Curado ◽

Sabhya Rana ◽

Ethan S. Benevides ◽

...

Keyword(s):

Gene Therapy ◽

Gene Delivery ◽

Motor System ◽

Viral Vectors ◽

Muscle Activation ◽

Neuromuscular Disorders ◽

Preclinical Studies ◽

Adeno Associated Virus ◽

Tongue Muscle ◽

Reduced Activity

AbstractDysfunction and/or reduced activity in the tongue muscles contributes to conditions such as dysphagia, dysarthria, and sleep disordered breathing. Current treatments are often inadequate, and the tongue is a readily accessible target for therapeutic gene delivery. In this regard, gene therapy specifically targeting the tongue motor system offers two general strategies for treating lingual disorders. First, correcting tongue myofiber and/or hypoglossal (XII) motoneuron pathology in genetic neuromuscular disorders may be readily achieved by intralingual delivery of viral vectors. The retrograde movement of viral vectors such as adeno-associated virus (AAV) enables targeted distribution to XII motoneurons via intralingual viral delivery. Second, conditions with impaired or reduced tongue muscle activation can potentially be treated using viral-driven chemo- or optogenetic approaches to activate or inhibit XII motoneurons and/or tongue myofibers. Further considerations that are highly relevant to lingual gene therapy include (1) the diversity of the motoneurons which control the tongue, (2) the patterns of XII nerve branching, and (3) the complexity of tongue muscle anatomy and biomechanics. Preclinical studies show considerable promise for lingual directed gene therapy in neuromuscular disease, but the potential of such approaches is largely untapped.

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Characterization of the Adeno-Associated Virus 1 and 6 Sialic Acid Binding Site

Journal of Virology ◽

10.1128/jvi.00161-16 ◽

2016 ◽

Vol 90 (11) ◽

pp. 5219-5230 ◽

Cited By ~ 35

Author(s):

Lin-Ya Huang ◽

Ami Patel ◽

Robert Ng ◽

Edward Blake Miller ◽

Sujata Halder ◽

...

Keyword(s):

Gene Therapy ◽

Sialic Acid ◽

Gene Delivery ◽

Binding Site ◽

Transduction Efficiency ◽

Cell Binding ◽

Adeno Associated Virus ◽

X Ray ◽

X Ray Crystallography ◽

Therapy Treatment

ABSTRACTThe adeno-associated viruses (AAVs), which are being developed as gene delivery vectors, display differential cell surface glycan binding and subsequent tissue tropisms. For AAV serotype 1 (AAV1), the first viral vector approved as a gene therapy treatment, and its closely related AAV6, sialic acid (SIA) serves as their primary cellular surface receptor. Toward characterizing the SIA binding site(s), the structure of the AAV1-SIA complex was determined by X-ray crystallography to 3.0 Å. Density consistent with SIA was observed in a pocket located at the base of capsid protrusions surrounding icosahedral 3-fold axes. Site-directed mutagenesis substitution of the amino acids forming this pocket with structurally equivalent residues from AAV2, a heparan sulfate binding serotype, followed by cell binding and transduction assays, further mapped the critical residues conferring SIA binding to AAV1 and AAV6. For both viruses five of the six binding pocket residues mutated (N447S, V473D, N500E, T502S, and W503A) abolished SIA binding, whereas S472R increased binding. All six mutations abolished or decreased transduction by at least 50% in AAV1. Surprisingly, the T502S substitution did not affect transduction efficiency of wild-type AAV6. Furthermore, three of the AAV1 SIA binding site mutants—S472R, V473D, and N500E—escaped recognition by the anti-AAV1 capsid antibody ADK1a. These observations demonstrate that common key capsid surface residues dictate both virus binding and entry processes, as well as antigenic reactivity. This study identifies an important functional capsid surface “hot spot” dictating receptor attachment, transduction efficiency, and antigenicity which could prove useful for vector engineering.IMPORTANCEThe adeno-associated virus (AAV) vector gene delivery system has shown promise in several clinical trials and an AAV1-based vector has been approved as the first gene therapy treatment. However, limitations still exist with respect to transduction efficiency and the detrimental effects of preexisting host antibodies. This study aimed to identify key capsid regions which can be engineered to overcome these limitations. A sialic glycan receptor recognition pocket was identified in AAV1 and its closely related AAV6, using X-ray crystallography. The site was confirmed by mutagenesis followed by cell binding and transduction assays. Significantly, residues controlling gene expression efficiency, as well as antibody escape variants, were also identified. This study thus provides, at the amino acid level, information for rational structural engineering of AAV vectors with improved therapeutic efficacy.

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Graphene oxide–hydroxyapatite nanocomposites effectively deliver HSV-TK suicide gene to inhibit human breast cancer growth

Journal of Biomaterials Applications ◽

10.1177/0885328218788242 ◽

2018 ◽

Vol 33 (2) ◽

pp. 216-226 ◽

Cited By ~ 4

Author(s):

Tuck-Yun Cheang ◽

Yi-Yan Lei ◽

Zhan-Qiang Zhang ◽

Hong-Yan Zhou ◽

Run-Yi Ye ◽

...

Keyword(s):

Gene Therapy ◽

Graphene Oxide ◽

Gene Delivery ◽

Cancer Cells ◽

Transfection Efficiency ◽

Suicide Gene ◽

Suicide Gene Therapy ◽

Human Breast ◽

Kinase Gene ◽

Ganciclovir Treatment

Gene therapy with herpes simplex virus thymidine kinase gene (HSV-TK), which is also known as “suicide” gene therapy, is effective in various tumor models. The lack of a safe and efficient gene delivery system has become a major obstacle to “suicide” gene therapy. In this study, the cytotoxicity and transfection efficiency of graphene oxide–hydroxyapatite (GO–Hap) were analyzed by MTS and flow cytometry, respectively. A series of assays were performed to evaluate the effects of GO–HAp/p-HRE/ERE-Sur-TK combined with ganciclovir treatment on growth of human breast normal and cancer cells. The results showed that GO–HAp nanocomposites effectively transfected cells with minimum toxicity. GO–HAp/p-HRE/ERE-Sur-TK combined with ganciclovir treatment inhibited the proliferation and induced cell apoptosis in cancer cells, while the cytotoxic effects are tolerable in normal breast cells. We conclude that the GO–HAp nanocomposites have significant potential as a gene delivery vector for cancer therapy.

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