We discuss scenarios with wrong-sign (WS) Yukawa couplings for the discovered Higgs boson in the Yukawa-aligned two Higgs doublet model. In the WS scenario, Yukawa couplings for down-type quarks and/or charged leptons have an opposite sign as compared to those of the Higgs boson in the standard model, which can be consistent with current flavor data and the Higgs signal strengths. The phenomenology of additional Higgs bosons in such a scenario can be significantly different from that with right-sign Yukawa couplings, mainly due to a larger Higgs boson mixing to be required in the wrong-sign case. We show the parameter space which is excluded or explored by direct searches for the additional Higgs bosons at the current and high-luminosity LHC under the constraints from perturbative unitarity and vacuum stability. In particular, we find that most of the parameter space is explored in the WS scenario with the Type-X (lepton specific) Yukawa interaction which is a special case of the Yukawa alignment realized by imposing a softly-broken ℤ2 symmetry. We propose that multi-Higgs events from pair productions of the additional Higgs bosons can be the smoking gun signature to probe the WS scenario, and give the expected number of events at the high-luminosity LHC.