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The Trinational Metropolitan Region Upper Rhine (TMO33 Official German abbreviation of the region, referring to Trinationale Metropolregion Oberrhein.) is a polycentric and highly urbanized region in Central Europe. The impacts of climate change onto the regional enterprises are complex and differ sectorwise and according to geographical factors. In this paper, we take a differentiated look at climate change impacts and adaptation efforts in nine of the region’s most affected sectors: agriculture, forestry & wood processing, food & beverage industries, water supply, logistics, winter tourism, manufacturing industries and the building sector. Framed by risk concepts we evaluate 157 interviews with company representatives and an additional online survey (n = 100) among small and medium-sized enterprises. The interviews and the online survey were carried out within the Interreg projects Clim’Ability and Clim’Ability Design during the years 2016–2021. We present an overview of sector-specific patterns of climate change impacts and adaptation efforts. Furthermore, we investigate the affected business units and fields of action for adaptation measures in a cross-sector perspective. Our results show that heat is the most relevant climatic stressor for a majority of the regional enterprises, followed by general warming and droughts. Extreme weather events like heavy rain, floods, thunderstorms or hail also play a considerable role, but adaptation to those phenomena appears to be less developed. This is an important starting point for future adaptation strategies and sector-specific recommendations for policy makers and local stakeholders.