This effect is unlikely to be caused by off-target impact: in virgin mice, even four doses of C188-9 did not result in a significant decrease of pSTAT5+ cells, increase of apoptosis, or decrease of size in early lesions (p=0.6, 0.8, and 0.7, respectively; n = 3 mice; data not shown).
To determine if the increase in LC3-II was due to inhibition of LRRK2 kinase activity or to an off target impact of LRRK2-in1, a dose response with a structurally distinct inhibitor of LRRK2 kinase activity, CZC-25146 (Supplemental Fig. S2A), was carried out [27].
Toxicity considerations are particularly critical in this context to minimize potential off-target phenotypic impacts on outcome measures.
Two patients in our study experienced CVA, and although these events were likely unrelated to the study drugs and neither patients experienced hypertension, the incidence of hypertension in our study raises the question of whether this combination may have off-target effects impacting vascular endothelium.
Broad-spectrum agents are indeed pleiotropic and, among various "off-target effects" can impact immune system function.
This transient impact on M45-specific CD8+ T cells could be due to a release of immunological pressure on the virus allowing viral transcription to proceed until the next immunological checkpoint is reached [ 13], or could be due to "off-target" or nonspecific impacts of the therapy.
However, inS3-54 may have off-target effects that can impact on cell survival, making it more effective in suppressing cancer cell survival.
Small interfering RNAs can trigger unintended, microRNA-like off-target effects, but the impact of these effects on functional studies has been controversial.
Finally, a caveat to consider with a small molecule like dorsomorphin is the potential impact on off-target effects.
Furthermore, the ability to probe a given target with multiple siRNA duplexes should decrease the impact of off-target effects on hit identification, if one sets a threshold that a minimum of two non-overlapping siRNA duplexes should yield an overlapping phenotype [11].
However, in preparing bisulfite libraries for sequencing, the use of ExoSAP-IT is not recommended due to potential off-target nuclease effects which may impact downstream methylation analysis.
