Novel use of FDA-approved drugs identified by cluster analysis of behavioral profiles

Abstract

Repurposing FDA-approved drugs is an efficient and cost-effective approach in the development of therapeutics for a broad range of diseases. However, prediction of function can be challenging, especially in the brain. We screened a small-molecule library with FDA-approved drugs for effects on behavior. The studies were carried out using zebrafish larvae, imaged in a 384-well format. We found that various drugs affect activity, habituation, startle responses, excitability, and optomotor responses. The changes in behavior were organized in behavioral profiles, which were examined by hierarchical cluster analysis. One of the identified clusters includes the calcineurin inhibitors cyclosporine (CsA) and tacrolimus (FK506), which are immunosuppressants and potential therapeutics in the prevention of Alzheimer’s disease. The calcineurin inhibitors form a functional cluster with seemingly unrelated drugs, including bromocriptine, tetrabenazine, rosiglitazone, nebivolol, sorafenib, cabozantinib, tamoxifen, meclizine, and salmeterol. We propose that drugs with ‘CsA-type’ behavioral profiles are promising candidates for the prevention and treatment of Alzheimer’s disease.

Modulation of calcineurin signaling during development

“…Other laboratories have previously shown that zebrafish larvae initially startle in response to sound pulses with a 1‐s interval, but quickly habituate to such stimuli (Best et al., 2008; Wolman et al., 2011). Subsequent studies in our own laboratory showed that habituation to sound stimuli with a 1‐s interval can be measured in a 384‐well format (Tucker Edmister, Del Rosario Hernandez, et al., 2022; Tucker Edmister, Ibrahim, et al., 2022). CsA and FK506 were used to suppress calcineurin signaling, proINDY was used to activate calcineurin signaling, and cotreatments were carried out to examine if proINDY is an effective therapeutic to restore calcineurin signaling.…”

“…We previously found that acute CsA and FK506 treatments induce hyperactivity in response to acoustic stimuli with a 1‐s interval, a behavior that we refer to as excitability (Tucker Edmister, Del Rosario Hernandez, et al., 2022; Tucker Edmister, Ibrahim, et al., 2022). In the present study, we examined effects of developmental exposures on excitability (Figure 4(c)).…”

“…Zebrafish larvae are known to swim in the same direction as a series of moving lines, which is referred to as an OMR (Naumann et al., 2016; Neuhauss, 2003; Portugues & Engert, 2009). We previously found that such behaviors can be imaged in four 96‐well plates for automated analyses of OMRs in a 384‐well format (Thorn et al., 2019; Tucker Edmister, Del Rosario Hernandez, et al., 2022; Tucker Edmister, Ibrahim, et al., 2022). Zebrafish have full‐color vision and OMRs can be measured using red, green, and blue lines (Tucker Edmister, Del Rosario Hernandez, et al., 2022; Tucker Edmister, Ibrahim, et al., 2022).…”

“…Zebrafish embryos and larvae were treated with 10 μM CsA (Enzo Life Sciences), 1 μM FK506 (Enzo Life Sciences), 1 μM rapamycin (Santa Cruz Biotechnology), 5 μM proINDY (Tocris Bioscience), or cotreatments of either 10 μM CsA + 5 μM proINDY or 1 μM FK506 + 5 μM proINDY. Prior studies in zebrafish have shown that these concentrations did not affect larval survival, but did induce changes in larval behavior (Clift et al., 2015; Tucker Edmister et al., 2022). Similarly, a 10 μM CsA exposure from 1 to 4 dpf did not affect survival or body length in zebrafish larvae (Robinson et al., 2017).…”

“…These studies may be expanded using other modulators of calcineurin signaling. For example, we recently found several US Food and Drug Administration‐approved compounds with CsA‐type and INDY‐type behavioral profiles (Tucker Edmister, Del Rosario Hernandez, et al., 2022). In addition, zebrafish lines that express the calcium indicator GCaMP can be used to image neural activity in the brain (Muto et al., 2013), bridging the gap between drug treatments and behavior.…”