Multispecies transport shapes electrochemical environments in perforated cells

Cells maintain asymmetric electrochemical environments across selectively permeable plasma membranes. Ion channels on plasma membranes redistribute small ions, whereas large openings such as gasdermin pores allow both ions and proteins to cross the membrane, generating a multispecies transport regime. The transport of charged ions and proteins shapes electrochemical environments, leading to osmolarity changes, which in turn promotes water influx and cell swelling. Although biologists have observed the phenotypic progression from pore formation to cell swelling for decades, the molecular transport events underlying this progression remain elusive. To elucidate how pore formation drives downstream cellular effects, we establish quantitative multispecies transport frameworks to investigate gasdermin-perforated cells. We start by developing a reductionist model and benchmarking it against analytical Donnan predictions. We then explore three-dimensional geometries containing individual pores or pore clusters that form either simultaneously or sequentially. We reveal that ion-protein timescale separation generates non-canonical Donnan-like states, pore and cargo properties tune local transport, pore clustering modulates osmotic gradients, and sequential pore formation produces feedback-dependent dynamics. This work deepens our mechanistic understanding of cellular responses to membrane damage and informs the therapeutic design of pore-forming proteins with tunable cellular effects.