Current Projects

 

Gene Therapy Vector Platforms

M13 iPhAGE Platform Optimization

The iPhAGE system is an advanced, bacteriophage-based gene delivery platform engineered to overcome critical limitations in current gene therapy, including safety, efficiency, and production challenges. iPhAGE ensures the targeted delivery of highly pure, minimized DNA vectors, free from immunogenic bacterial elements. It shows enhanced gene expression and targeted cellular uptake.

We continue to optimize and improve our scalable, cost-effective platform to further enhance its efficiency, efficacy, and broad therapeutic potential across numerous applications.

Novel Anellovirus-Based Vector Biomanufacturing Production Platform

This is a novel, highly efficient, and cost-effective production platform for anellovirus-based gene therapy vectors. Utilizing an E. coli-based production system, this system enables the scalable production of highly pure, circular ssDNA vectors encapsidated within anellovirus capsid proteins, termed “anellomids.” Anellomids provide superior gene expression and potential redosability, and effectively addresses critical gene therapy limitations, including safety concerns, manufacturing complexity, and suboptimal transgene expression, thus positioning them as a promising solution for broad therapeutic applications.

Novel Adeno-Associated Virus Biomanufacturing Production Platform

Adeno-associated virus (AAV) is an unenveloped virus known for its ability to effectively and securely transport genetic material into cells, making it valuable for gene therapy. Although industrial manufacturing methods for AAV vector production have advanced, challenges remain in terms of quality and production costs, hindering wider development.

The limited scalability of current methods has resulted in prohibitively high costs for many patients seeking AAV gene therapy. We seek to develop a novel AAV vector production method to mitigate these issues.


Applications

Targeted Bacteriophage Gene Delivery to Astrocytes

The filamentous bacteriophage M13 is a single-stranded DNA phage that co-exists with its Escherichia coli host after infection. M13 possesses several attractive characteristics for gene delivery and have a well-documented ability to cross the blood-brain barrier which is an exploitable trait that can be applied to targeted phage delivery to the CNS. Astrocytes have been shown to differentiate into functional neurons. This ability makes them an ideal target for gene delivery and cell transformation.

The goal of this project is to show that intravenous administration of M13 miniphagemids, engineered to carry LCC DNA vectors (iPhAGEs) will effectively cross the blood brain barrier and allow for the reprogramming of astrocytes.

Redosable Gene Delivery Platforms for Retinal Applications

Stargardt Disease, an inherited disease characterized by progressive vision loss, is driven by mutations in the large ABCA4 gene, exceeding the capacity of traditional gene therapy vectors. Our research addresses this with "ministring DNA" (msDNA) minivectors that efficiently deliver the therapeutic ABCA4 gene to the retina.

To further enhance delivery and redosability, we are developing an iPhAGE system utilizing engineered M13 miniphagemids. These phage-based vectors are designed for targeted delivery to photoreceptors offering a promising path for effective and personalized treatment of Stargardt Disease and other ocular disorders.

 
 

The Effects of Amyloid-Beta on Neuronal Transcription

Alzheimer's disease (AD) is a neurodegenerative disorder marked by cognitive decline. The goal of this project was to demonstrate that M13-based miniphagemid vectors could be engineered for targeted gene delivery to neurons, laying foundational groundwork for precise therapeutic interventions in Alzheimer's disease.

M13 miniphagemids were engineered to display Angiotensin IV for neuronal targeting, leveraging its receptor's role in brain delivery. Their genome was also engineered to encode a neural specific promoter, Synapsin 1, for targeted expression, as well as carried a therapeutic gene, ZIP3, or reporter gene, eGFP.

Targeted Miniphagemid Mediated Anti-Angiogenic VLPs

Molecular targeted therapy has emerged as a promising strategy to treat cancer over the last several years. Unlike the broad-spectrum cytotoxic drugs prescribed for conventional chemotherapy, targeted therapy is designed to address specific molecular changes which are unique to a specific cancer type. We developed a system for targeted miniphagemid-mediated delivery of DNA encoding VLPs displaying anti-tumour peptides to tumour cells. We hypothesized that this would then induce the tumor cells to produce HPV VLPs displaying VGB4.

 

Lambda Phage & Vaccines

Identification of E. coli Host Genes Influencing T4rII Exclusion Phenotype

The rex genes (rexA, rexB) of bacteriophage Lambda (λ) are able to prevent mutant bacteriophage T4rII plaque formation of E. coli hosts lysogenized by λ. The expression of these two genes of the Rex system is primarily regulated by the repressor gene cI from the PM promoter.

This project aims to isolate and identify relevant host mutations that could influence the Rex phenotype. Through this work, the manifestation of Rex has been linked to genes underlying key host stress responses.

Universal Virus-Like Particle for Coronavirus

The human immune system, while capable of generating neutralizing antibodies against pathogens, faces a constant challenge from evolving virus populations, like COVID-19, which generate new strains to evade existing immunity. To address this, we are developing a Universal Virus-Like Particle (UVLP), leveraging a "consensus sequence" derived by averaging genes from closely related coronaviruses. The UVLP is designed to induce a broadly neutralizing antibody response in order to offer durable protection against not just current COVID-19 variants, but also future mutations and emerging coronaviruses.


A selection of associated publications:


Research Articles

  1. A novel miniaturized filamentous phagemid as a gene delivery vehicle to target mammalian cells. Read here.

  2. An E. coli-based platform for the production and assembly of anellovirus vectors.

    Read here.

  3. Construction and characterization of a novel miniaturized filamentous phagemid for targeted mammalian gene transfer. Read here.

  4. The effects of amyloid beta aggregation on neuronal transcription. Read here.

  5. EGFR-targeted bacteriophage lambda penetrates model stromal and colorectal carcinoma tissues, is taken up into carcinoma cells, and interferes with 3-dimensional tumor formation. Read here.

  6. A snapshot of the λ T4rII exclusion (Rex) phenotype in Escherichia coli. Read here.

  7. Identification of Escherichia coli Host Genes That Influence the Bacteriophage Lambda (λ) T4rII Exclusion (Rex) Phenotype. Read here.

  8. Lambda bacteriophage nanoparticles displaying GP2, a HER2/neu derived peptide, induce prophylactic and therapeutic activities against TUBO tumor model in mice. Read here.

  9. Immunogenicity and antitumor activity of the superlytic λF7 phage nanoparticles displaying a HER2/neu-derived peptide AE37 in a tumor model of BALB/c mice. Read here.

  10. Lambda Phage nanoparticles displaying HER2 derived E75 peptide induce effective E75-CD8+ T response. Read here.

  11. Construction and analysis of a genetically tuneable lytic phage display system. Read here.

  12. Graphical analysis of flowcytometer data for characterizing controlled fluorescent protein display on Lamba phage. Read here.

Review Articles

  1. Advances in gene-based vaccine platforms to address the COVID-19 pandemic. Read here.

  2. Bacteriophage interactions with mammalian tissue: Therapeutic applications. Read here.

  3. The viral approach to breast cancer immunotherapy. Read here.

  4. Phage-based nanomedicines as new immune therapeutic agents for breast cancer. Read here.

  5. Bacteriophage lambda display systems: developments and applications. Read here.


Patents

  1. Nucleic Acid Delivery System. Find online.

  2. Vectors for Producing Virus-Like Particles and Uses Thereof. Find online.

  3. DNA Vector Production System. Find online.

  4. DNA Vector Production System. Find online.

  5. EGFR Binding Moiety-Presenting Bacteriophages for Tumor Treatment. Find online.

  6. Fusion Protein Comprising Bacteriophage  λ GPD  Fused to a GP2 Antigen Derived from HER-2/NEU. Find online.

  7. Peptide Displaying Bacteriophage Nanoparticles and Related Compositions and Methods. WO 2019/102265 A1. Find online.


Books

  1. Gene Delivery Imaging and Evaluation Systems. Find online.

  2. Bacteriophage Applications - Historical Perspective and Future Potential. Find online.