Supplementary data for: Comparison of transcriptomic profiles between HFPO-DA and prototypical PPARa, PPARg, and cytotoxic agents in wild-type and PPARa knockout mouse hepatocytes

Recent in vitro transcriptomic analyses for the short-chain
polyfluoroalkyl substance (PFAS), HFPO-DA (ammonium,
2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)-propanoate), support
conclusions from in vivo data that HFPO-DA-mediated liver effects in mice
are part of the early key events of the peroxisome proliferator-activated
receptor alpha (PPARa)activator-induced rodent hepatocarcinogenesis mode
of action (MOA). Transcriptomic responses in HFPO-DA-treated rodent
hepatocytes have high concordance with those treated with a PPARa agonist
and lack concordance with those treated with PPARg agonists or cytotoxic
agents. To elucidate whether HFPO-DA-mediated transcriptomic responses in
mouse liver are PPARa-dependent, additional transcriptomic analyses were
conducted on samples from primary PPARaknockout (KO) and wild-type (WT)
mouse hepatocytes exposed for 12, 24 or 72 hours with various
concentrations of HFPO-DA, or well-established agonists of PPARa (GW7647)
and PPARg (rosiglitazone), or cytotoxic agents (acetaminophen or
d-galactosamine). Pathway and predicted upstream regulator-level responses
were highly concordant between HFPO-DA and GW7647 in WT hepatocytes. A
similar pattern was observed in PPARa KO hepatocytes, albeit with a
distinct temporal and concentration-dependent delay potentially mediated
by compensatory responses. This delay was not observed in PPARa KO
hepatocytes exposed to rosiglitazone, acetaminophen, d-galactosamine. The
similarity in transcriptomic signaling between HFPO-DA and GW7647 in both
the presence and absence of PPARa in vitro indicates these compounds share
a common mechanism of action and supports the PPARa-dependence of
HFPO-DA-mediated effects in mouse liver.