Anti‐Methamphetamine Antibody Gene Therapy Protects Against Some METH‐Induced Inflammation
Authors: Bolden Chris , McLeroy‐Charles Kennede , Hay Charles E. , Ewing Laura E. , Margaritis Paris , Peterson Eric C. 
Published: June 2021

Methamphetamine (METH) is a highly abused, addictive drug that can adversely affect the peripheral immune response, causing immune dysregulation in the user. This can lead to an overproduction of cytokines, chemokines, and cellular adhesion molecules causing potential physiologic damage. Thus novel, sustained treatments that focus on mitigating these effects should be evaluated.

Our laboratory is combining an anti‐METH single chain variable antibody‐fusion fragment (7F9‐Fc) with adeno‐associated virus (AAV) particles as a potential gene therapy. AAV‐7F9‐Fc works as a pharmacokinetic antagonist to slow and reduce entry of METH into the brain and can achieve lasting serum concentrations in mice (>7 months) from a single treatment. Thus, this therapy has the clinical potential to attenuate the effects of periodic METH relapse during rehabilitation and possibly decrease associated physiological complications such as inflammation.

We hypothesized that an AAV‐delivered anti‐METH antibody fragment (AAV‐7F9‐Fc) will provide protection from METH‐induced inflammation. To test this hypothesis, we established a mouse model of METH‐induced early withdrawal inflammation in our laboratory. Male BALB/c mice were divided into six groups (n=5/group); 1) Saline + Saline, 2) Saline + METH, 3) LPS, 4) AAV‐7F9‐Fc + Saline, 5) AAV‐7F9‐Fc + METH, 6) AAV‐empty vector + saline. METH administration followed a dosing regimen of 1 mg/kg i.p. for seven consecutive days with sacrifice after 72 hrs. We performed a MSD Proinflammatory Panel multiplex analysis to measure important serum cytokine and chemokine levels (e.g., IL‐10, IL‐6, IL‐1®). In brain tissues, we measured markers of glial cell activation, glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule (Iba1) and proinflammatory cytokines via qPCR.

Our results indicate that this dosing regimen lead to a significant (p < 0.05) increase in proinflammatory mediators (e.g., IL‐1® and IL‐6) in the striatum of Saline + METH mice, but not AAV‐7F9‐Fc + METH mice. In the hippocampus and frontal cortex, the Saline + METH mice showed a significant increase in the mRNA transcripts of IL‐6 & IL‐1® vs those mice treated with AAV‐7F9‐Fc + METH. Noticeably, GFAP was also increased in all observed brain regions of Saline + METH vs those treated with AAV‐7F9‐Fc + METH. Our results also indicate that this dosing regimen lead to a significant increase in serum proinflammatory cytokines (e.g., IL‐1® and IL‐6) vs the AAV‐7F9‐Fc + METH mice. Taken together, these data indicate that a chronic, low dose METH regimen can induce proinflammatory cytokine elevations in serum and brain, increase activation of glial cell markers in brain tissue, and that AAV‐7F9‐Fc treatment has the potential to mitigate some of these proinflammatory effects.

Support or Funding Information

Funded by NIH R01‐DA036600, R01 DA036600‐03S1, T32‐GM106999, and R25GM083297

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.