The parents of Niccolò and his brother Francesco I Gattilusio are not known, although based on the heraldic evidence of their inscriptions, Anthony Luttrell argues that their mother was a member of the Doria family.
As the former was rewarded by the Byzantine emperor John V Palaiologos with the island of Lesbos, so Niccolò received the coastal city of Ainos, which he took title to at some point between 1376 and 1379.
[1] From 1384 to 1387 he served as regent for his nephew Francesco II Gattilusio until the two fell out in an argument between them.
[2] When his nephew died in an unusual accident, Niccolò served once more as regent, this time for Francesco's son Jacopo.
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