We review the current understanding of heavy quark parton distributions in nucleons and their impact on deep inelastic scattering, collider physics, and other processes at high energies. The determination of the heavy-quark parton distribution functions is particularly significant for the analysis of hard processes at LHC energies, including the forward rapidity high xF domain. The contribution of “intrinsic” heavy quarks, which are multiply connected to the valence quarks of nucleons, is reviewed within non-perturbative physics which provides new information on the fundamental structure of hadrons in QCD. A new prediction for the non-perturbative intrinsic charm–anticharm asymmetry of the proton eigenstate has recently been obtained from a QCD lattice gauge theory calculation of the proton’s GEp(Q2) form factor (Sufian et al., 2020). This form factor only arises from non-valence quarks and anti-quarks if they have different contributions in the proton’s eigenstate. This result, together with the exclusive and inclusive connection and analytic constraints on the form of hadronic structure functions from Light-Front Holographic QCD (LFHQCD) predicts a significant non-perturbative c(x,Q)−c̄(x,Q) asymmetry in the proton structure function at high x, consistent with the dynamics predicted by intrinsic charm models. Recent ATLAS data on the associated production of prompt photons and charm-quark jets in pp collisions at s=8TeV has provided new constraints on non-perturbative intrinsic charm and tests of the LGTH predictions. We also focus on other experimental observables which have high sensitivity to the intrinsic heavy contributions to PDFs.