Integrated and transdisciplinary approaches are necessary in hotspots research where the intention is to influence policy and practice. Knowing that climate change will impact major ecosystem services and the sustainability of life support systems, a critical examination of the hotspot concept and approach is undertaken to pursue synergistic responses. Hotspots 2.0 embodies current thinking about planning towards multiple drivers of change and seeing human and natural systems as mutually inter-dependent and benefiting from integrated policy approaches. Such proposed adaptation interventions to inter-related stressors will complement biodiversity conservation, disaster risk reduction, and human wellbeing. Through a systematic review, we assess 114 relevant peer review cases to examine integrative responses to climatic and non-climatic vulnerabilities in various hotspot regions. Furthermore, we illustrate the utility of the Hotspots 2.0 approach using emerging insights from the ‘Collaborative Adaptation Research Initiative in Africa and Asia’ in semi-arid regions, deltas, and glacier-fed river basin hotspots.