Electrically regulated cell‐based intervention for viral infections

Abstract

This work reports on an engineered cell that—when electrically stimulated—synthesizes a desired protein, that is, ES‐Biofactory. The platform has been used to express interferon (IFN)‐β as a universal antiviral protein. Compelling evidence indicates the inevitability of new pandemics and drives the need for a pan‐viral intervention that may be quickly deployed while more specific vaccines are in development. Toward this goal, a fast‐growing mammalian cell (Chassis) has been engineered with multiple synthetic elements. These include—(1) a voltage‐gated Ca2+ channel (Voltage‐Sensor) that, upon sensing the electric field, activates the (2) Ca2+‐mediated signaling pathway (Actuator) to upregulate (3) IFN‐β, via an engineered antiviral transgene (Effector), that is, ES‐Biofactory➔IFN‐β. The antiviral effects of the ES‐Biofactory➔IFN‐β have been validated on severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2)‐infected cells. The irradiated ES‐Biofactory, that does not exhibit oncogenic capacity, continues to exert antiviral effect. The resulting ES‐Biofactory➔IFN‐β uses a novel signaling pathway that, unlike the natural IFN synthesis pathway, is not subject to viral interference. Once clinically validated, the ES‐Biofactory will be a universal antiviral cell therapy that can be immediately deployed in the event of an outbreak. The platform may also be useful in treating other diseases including cancer and autoimmune disorders.

Primary T‐cell‐based delivery platform for in vivo synthesis of engineered proteins

“…As a result, this platform technology opens new horizons for treating a variety of diseases. For example, we have already employed immortalized cell lines to establish the feasibility of using the synthetic pathway for localized delivery of various type‐I and type‐III interferons for targeting SARS‐CoV‐2 22,24 . Our current efforts are geared toward applying our antigen‐specific primary T‐cell‐based system for in situ synthesis of such biologic drugs to exert their therapeutic effects within the local tumor microenvironment.…”

“…For example, we have already employed immortalized cell lines to establish the feasibility of using the synthetic pathway for localized delivery of various type‐I and type‐III interferons for targeting SARS‐CoV‐2. 22 , 24 Our current efforts are geared toward applying our antigen‐specific primary T‐cell‐based system for in situ synthesis of such biologic drugs to exert their therapeutic effects within the local tumor microenvironment. Our parallel efforts, in continuation of this work, also include improving our delivery system so as to increase the amount of protein‐based drugs produced from a single cell while reducing its expression that may occur under unstimulated conditions.…”

“… 21 In this work, using a CAR with specificity for folate receptor alpha (FRα) and a reporter enzyme to represent the desired biologic, we described a simple process to efficiently scale the primary T‐cell‐based delivery platform (Figure 1 ). As recently demonstrated by us, 13 , 14 , 22 , 23 , 24 , 25 however, the DNA template of the antigen‐sensing scFv domain and the reporter enzyme can be exchanged to redirect the specificity of this platform to another clinically relevant antigen and express a protein for sensing 23 , 25 or therapeutic 22 , 24 function.…”

Genetically engineered pair of cells for serological testing and its application forSARS‐CoV‐2

“…(3) a practical approach for manufacturability; and (4) due to its modularity, 7,8 this cell-based testing platform that can be transformed into a therapeutic tool by exchanging the Reporter protein with a therapeutic protein, as demonstrated by our group. 22,23 This work complements existing serological testing technologies 18,20,24,25 enabling informed public health decisions required to reopen economies. For example, it will enable identification of individuals who may have seroconverted due to vaccination or prior SARS-CoV-2 infection.…”

“…Our data show that the performance characteristics of the DxCell‐Complex serology test could exceed the minimum requirements listed for serology tests undergoing FDA emergency use authorization submission 21 . The DxCell‐Complex has the potential to identify individuals with recent or prior SARS‐CoV‐2 infections, and offers unique advantages by enabling: (1) cost‐effective mass‐testing capabilities in decentralized healthcare facilities and with easy sample preparation steps; (2) an easily transportable quantitative, self‐replicating diagnostic tool that works in a label‐free manner without signal amplification steps; (3) a practical approach for manufacturability; and (4) due to its modularity, 7,8 this cell‐based testing platform that can be transformed into a therapeutic tool by exchanging the Reporter protein with a therapeutic protein, as demonstrated by our group 22,23 …”