A new bisdehydro[12]annulene derivative having a thiophene-fused structure has been synthesized. This highly twisted pi-conjugated macrocycle with two acetylene moieties in close proximity produces a [2+2]-type alkyne cycloadduct by either photoirradiation or mild heating without any transition metals. Theoretical calculations reveal that the thermal reaction proceeds through successive 8 pi and 4 pi electrocyclic reactions, while the photochemical reaction is an asynchronous concerted [2+2] cycloaddition. The fused structure with the less-aromatic thiophene ring is crucial for achieving this reaction. The cycloadduct, thiophene-fused biphenylene, has significant potential as a new polycyclic pi-scaffold for electronic applications.