Substantial SARS-CoV-2 spread over the last 5 years of the COVID-19 pandemic has enabled the continued accumulation of viral genomic changes. Enrichment in changes in the spike glycoprotein has enabled increased resistance to neutralising antibodies and concomitant changes to entry requirements that increased viral transmission fitness. The mechanistic basis of this increased transmission has yet to be fully resolved and ironically, genetic engineering of cell lines with prior entry factors angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) has led to conditions of viral attenuation following the appearance of Omicron. Herein, we demonstrate the evolving use of ACE2/TMPRSS2 dependency over the last five years of the pandemic. Mechanistically we show a continual shift in tropism that relates to the dynamics of its ACE2 and TMPRSS2 at three levels. Firstly, SARS-CoV-2 over time has shifted from furin to TMPRSS2 in enabling S1/S2 Spike cleavage. Secondly, this cleavage by TMPRSS2 is negatively regulated by association of TMPRSS2 with the ACE2 Collectrin-Like Domain (CLD). Finally, the oligomerization of ACE2 with either solute carrier, SLC6A19 or SLC6A20, at its CLD positively regulates TMPRSS2 cleavage of S1/S2 through limiting ACE2-TMPRSS2 complexes. To conclude our data herein observe the evolution of SARS-CoV-2 tropism towards the use of ACE2 pools primarily associated with SLC6A19/SLC6A20 which enables TMPRSS2 usage to sustain high levels of onward transmission.