64Cu-ATSM accumulates in tumor regions with upregulation of DNA repair.
64Cu-ATSM high-uptake regions show enrichment of CD133+ cancer stem-like cells.
Nucleic acid (NA) incorporation is facilitated in 64Cu-ATSM high-uptake regions.
64Cu-ATSM therapy with an NA antimetabolite synergistically inhibited tumor growth.
The sensitized 64Cu-ATSM therapy effectively reduced %CD133+ cells.