A new well-dated (18 14C dates on short-lived terrestrial plant macrofossils, 210Pb and 137Cs dating) fine-resolution palaeoecological sequence (pollen, coprophilous fungi, conifer stomata, macrofossils, charcoal) from Lago de Isoba (1400 m a.s.l.) spanning the past c. 17,000 years provides novel insights into the long-term dynamics of the high-elevation Cantabrian landscape in response to climate variability and human activities. Vegetation changes were mainly climate-driven during the Late-glacial and the Early Holocene. Thus, steppe-like environments prevailed during the Greenland Stadial 2.1a (GS-2.1a ≈ ‘Oldest Dryas’). Warmer and moister conditions during the Greenland Interstadial 1 (GI-1 ≈ ’Bølling-Allerød’) enabled successive expansions of Juniperus, Betula and Pinus. Steppe-like vegetation re-expanded during the cooler and drier Greenland Stadial 1 (GS-1 ≈ ‘Younger Dryas’). Afterwards, the Early Holocene warming triggered the rapid spread of mixed pine-broad-leaved deciduous mountain forests. Pollen, stomata and macrofossils indicate continuous, long-lasting (c. 10,850–1250 cal yr BP) local occurrence of Pinus sylvestris stands. The records of Cerealia and Plantago lanceolata pollen types (first Holocene occurrences dated at c. 5100 cal yr BP) evidence increased land use during the Late Holocene, which triggered substantial changes in forest ecosystems. Following a phase of intensified arable and pastoral farming, which involved the use of fire to create and maintain forest clearings, pinewoods expanded and reached their maximum development between c. 3600 and 3300 cal yr BP. Pinus probably outcompeted temporarily other woody taxa thanks to its resilience to low-intensity disturbances. Later, pinewoods underwent a stepwise, human-induced decline ending in their final massive collapse after c. 1150 cal yr BP. The highest intensity of agricultural and pastoral farming took place during the Middle Ages, following maximum fire occurrence. We conclude that the present-day open landscape has been shaped by millennia of land use that partly blurred the impact of climate variability.