Expert Consultation: Shaping Our Project

In order to gain some more insight into the nature of our project and its potential applications, we consulted two professors here at Westlake. First was Dr. Dijin Xu, who studies host-pathogen interactions and is focused on characterizing the different immune pathways and reaction mechanisms. Secondly is Dr. Dapeng Li, whose lab studies not only viral life cycles, but also the development of antiviral therapeutics and vaccine treatments. By talking to these two experts, we hoped that their knowledge would help us to more accurately assess the ability of our project to serve as an effective treatment.

Here we summarize key insights from consultations with immunology experts, Dr. Li and Dr. Xu, on the development of a novel broad-spectrum antiviral gene therapy. The discussion is structured around the AREA framework (Acknowledge, Research, Effect, Adjust), highlighting the cyclical process of addressing core challenges in the project.

Cycle 1: Target Identification and Therapeutic Scope

  • Acknowledge (The Problem): The primary hurdle in creating a "perfect" broad-spectrum antiviral is the immense diversity of viruses. A therapeutic that targets a single, highly specific viral protein is unlikely to be effective across multiple virus families.
  • Research (The Exchange): Consultation with Dr. Li emphasized the need to shift focus from individual viruses to conserved elements. The recommended research direction involves using proteomic and high-throughput screening technologies to systematically analyze viral life cycles. The goal is to identify common, essential steps—particularly in viral entry—that could serve as universal intervention points.
  • Effect (The Impact): This guidance broadened the project's scope from a virus-centered approach to a pathway-centered one. It established the core hypothesis: targeting a conserved host-pathogen interaction or a common step in the viral life cycle is the most viable strategy for broad-spectrum efficacy.
  • Adjust (The Refinement): The design of the project was adjusted to prioritize the identification of highly conserved viral entry mechanisms. However, this new focus immediately revealed a subsequent challenge: how to design a therapeutic that can act rapidly enough to block this narrow entry window.

Cycle 2: Therapeutic Design and Practical Application

  • Acknowledge (The New Problem): A therapy targeting viral entry is only feasible if it can be activated with high speed and efficiency. Furthermore, the chosen gene activation system itself must be both safe for human cells in vitro and suitable for delivery in vivo.
  • Research (The Exchange): Dr. Li stressed that translational feasibility must be a primary design consideration. Key questions were raised regarding the kinetics of the proposed gene activation system: its activation speed, its potential cytotoxicity over time, and the availability of safer, more efficient delivery mechanisms. Dr. Li noted that when given the opportunity to simplify a part, you should usually take it.
  • Effect (The Impact): This exchange forced a critical evaluation of our proposed technology's real-world applicability. The project's success criteria were expanded to include not just biological efficacy, but also pharmacokinetic and safety parameters. A system that takes too long to induce is useless, as is one that leaves too soon. We had to find a balance between the two.
  • Adjust (The Refinement): The project design was adjusted by reworking the delivery of each component in a seperate vector to improve both speed and safety. This way, the size of the pDNA wouldn't impede delivery, but we could still control rtTA and dCas9 under two different promoters.

Cycle 3: Balancing Efficacy and Safety in Gene Selection

  • Acknowledge (The New Problem): The strategy of enhancing innate immunity by upregulating antiviral genes introduces a significant safety risk: the potential for excessive inflammation and off-target cellular damage.
  • Research (The Exchange): Dr. Xu provided a critical framework for gene selection, outlining a key trade-off. Targeting upstream innate immune effectors (e.g., IFN signaling, cGAS-STING) provides broad antiviral effects but carries a high risk of triggering a damaging cytokine storm. Targeting downstream effectors is safer but may offer a narrower, less effective spectrum of antiviral activity. The emphasis was placed on a deep understanding of each protein's pleiotropic functions and the consequences of its long-term activation.
  • Effect (The Impact): This refined the project's target selection strategy from a simple search for "antiviral genes" to a nuanced balancing act between potency and safety. The functional characterization of any candidate gene must now include a thorough analysis of its role in inflammatory pathways.
  • Adjust (The Refinement): The project is now focused on identifying "sweet spot" targets, genes that are sufficiently downstream to minimize inflammatory risks but upstream enough to retain meaningful broad-spectrum activity. This has led to a renewed research phase, centered on mapping host-pathogen protein interactions to find critical, druggable nodes for intervention, as suggested by Dr. Xu, thereby initiating a new cycle of the AREA framework.