TNFα signaling results in RNA polymerase II-dependent transcriptional outcomes and downstream biological effects in human cardiomyocytes. TNFα signaling activates an NF-κB-mediated transcriptional program in AC16 cardiomyocytes. Integrated genomic analyses capture transcriptome alterations for all expressed Pol I, II, III transcripts in inflammatory cardiomyocytes. Pol II transcripts are the main drivers of the rapid and dynamic transcriptome changes, whereas non-Pol II transcripts show limited responses to TNFα . The Pol II dependent globally transcriptome changes include both protein coding and non-coding transcripts, which orchestrate the subsequent biological effects of inflammation on cardiomyocytes.