Resistance against all existing antimalarial drugs is becoming a major concern and highlights the need for the development of novel treatment strategies. Host-directed therapy (HDT) has potential for combatting malaria, and has the advantage of depriving the parasite of the most direct pathway to resistance, namely the selection of mutated targets under drug pressure. Previous studies have revealed that Plasmodium falciparum relies on the activation of host erythrocyte protein kinases for its own proliferation and survival. One host kinase in particular, the mitogen activation protein kinase kinase 1 or MEK1, has been shown to be activated in infected erythrocytes and hepatocytes. Trametinib is a highly selective MEK1 inhibitor that is approved for clinical use against melanoma. We showed that Trametinib inhibits parasite proliferation in vitro with low nanomolar potency, suggesting that MEK activation is required for parasite survival. Surprisingly, we were able to generate parasite lines that are resistant to Trametinib, showing a 30-fold increase in the IC50. Intriguingly, some of these parasites display dependency to the drug, and show optimal proliferation in the presence of approximately one micromolar Trametinib. Whole genome sequencing of clones obtained from these lines identified candidate mutations associated with the dependency phenotype. Our work provides novel insights into the molecular mechanisms of host-pathogen interactions between human erythrocytes and P. falciparum, and highlights human kinases as targets for the development of novel anti-malarials with an untapped mode of action.