Selective protection of human cardiomyocytes from anthracycline cardiotoxicity by small molecule inhibitors of MAP4K4

Abstract

Given the poor track record to date of animal models for creating cardioprotective drugs, human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have been proposed as a therapeutically relevant human platform to guide target validation and cardiac drug development. Mitogen-Activated Protein Kinase Kinase Kinase Kinase-4 (MAP4K4) is an "upstream" member of the MAPK superfamily that is implicated in human cardiac muscle cell death from oxidative stress, based on gene silencing and pharmacological inhibition in hPSC-CMs. A further role for MAP4K4 was proposed in heart muscle cell death triggered by cardiotoxic anti-cancer drugs, given its reported activation in failing human hearts with doxorubicin (DOX) cardiomyopathy, and its activation acutely by DOX in cultured cardiomyocytes. Here, we report successful protection from DOX in two independent hPSC-CM lines, using two potent, highly selective MAP4K4 inhibitors. The MAP4K4 inhibitors enhanced viability and reduced apoptosis at otherwise lethal concentrations of DOX, and preserved cardiomyocyte function, as measured by spontaneous calcium transients, at sub-maximal ones. Notably, in contrast, no intereference was seen in tumor cell killing, caspase activation, or mitochondrial membrane dissipation by DOX, in human cancer cell lines. Thus, MAP4K4 is a plausible, tractable, selective therapeutic target in DOX-induced human heart muscle cell death. With earlier diagnosis and improved treatments for cancer, cardiovascular disease has become the main alternative cause of death in cancer survivors 1-4. This heightened risk is ascribable to the cardiac toxicity of several routine anti-cancer agents including, especially, anthracyclines like doxorubicin (DOX) 5. Though some relief can result from standard heart failure medications, no approved therapy apart from the iron chelator dexrazoxane addresses the responsible cytotoxic mechanisms and effector pathways operating in the damaged cardiomyocytes. For anthracyclines, these include diverse reported mediators-binding to nuclear topoisomerase 2β, thereby triggering DNA double-strand breaks, p53-dependent apoptosis, mitochondrial dysfunction, reactive oxygen species, and programmed iron-dependent cell death (ferroptosis) 1,6,7. Cardiotoxicity can be acute, early, or late; early intervention based on subclinical abnormalities may be key to averting the delayed or cumulative effects 8. Notably, acute toxicity is seen in up to 30% of patients receiving anthracyclines, soon after infusion 4. Current cardioprotection trials in cancer rely chiefly on routine heart failure medications (ACE inhibitors, β-blockers, angiotensin receptor blockers), which mitigate the symptoms and signs, with little or no demonstrated impact on cardiomyocyte death. Therapeutic progress has been hampered, in part, by the failure of animal models alone to predict clinical success in cardioprotection. For instance, nearly all strategies for protection in ischemic heart disease, a more intensively studied indication, have failed between phases I and I...

Transcriptional co‐activators YAP1–TAZ of Hippo signalling in doxorubicin‐induced cardiomyopathy

“…Given that increased YAP might lead to drug resistance in cancer cells, 36 , 37 it is important to consider cardio‐selective overexpression when developing new cardioprotective strategies. 35 , 38 Cardiac‐specific YAP activation would selectively protect cardiac cells and allow doxorubicin to simultaneously exert its effect on cancer cells. 39 In turn, small‐molecule inhibitors of YAP/TAZ are considered potential therapies for various cancers, like mesothelioma and basal cell carcinoma.…”

Mitochondrial-Targeted Therapy for Doxorubicin-Induced Cardiotoxicity

“…Golforoush et al showed that the inhibition of the Mitogen-activated protein kinase kinase kinase kinase-4 (MAP4K4) can alleviate DOX-induced cardiotoxicity using both rat H9c2 cells and hPSC-cardiomyocytes [125]. MAP4K4 is an upstream member of the MAPK superfamily that can regulate cell death.…”

“…The same group has previously shown that inhibition of this pathway using shRNA and the small molecule inhibitor DMX-5804 can improve mitochondrial function, protect against lethal oxidative stress, and reduce cardiac ischemia reperfusion injury [137]. Golforoush et al [125] then proceeded to show that DMX-5804 could confer similar protection against DOX in in vitro models. In terms of mitochondrial homeostasis, Liang et al 2020 showed that liensinine, a newly identified inhibitor of mitophagy, can protect against DOX in in vitro and in vivo rodent models by inhibiting DRP1, a regulator of mitochondrial fission [126].…”

Nanomedicine in Oncocardiology: Contribution and Perspectives of Preclinical Studies

“…The evaluation of the cardiotoxicity of nanostructures can be performed both in vitro and in vivo. Human pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) emerge as a practical approach for the early in vitro screening of cardiotoxicity of new drugs (28)(29)(30). However, investigations on animal models are still required to develop their use in clinical practice despite some limitations.…”