One of the most applied tools for documenting cultural variability and tracing the cultural trajectories within the environmental context is Eco-Cultural Niche Modeling ECNM and their associated methodology. The ECNM has been adapted by archeologists and not only from the bio-computational architecture to explore the interactions between cultural and natural systems and to understand how ecological dynamics influenced adaptations and movements of prehistoric populations. Therefore, even if the concept of Ecological Niche Modeling ENM is derived from the Biology disciplines and commonly used in biodiversity or geographic ecology studies, the ECNM applies the same methodology to analyses archaeological record and prehistoric human culture via GIS softwarersquo;s. Also, the niche breadth characterization offers the opportunity to evaluate quantitatively the links between a given adaptive system and ecological constraints which provides valuable information for archaeology. In this work, ten independent climatic and topographic environmental variables were interpolated and ECNM techniques were used to determine whether these differences in the geographic distributions and niche breadth are consequences of differences for five Cucuteni-Trypillia groups which flourished in Eastern Europe during the Eneolithic cal. 5,400/5,300 - 2,800/2,700 BCE. The outcomes show that the eco-cultural niches of Cucuteni-Trypillia groups is significantly overlapping and the expansion trend of the cultural groups from Eastern Carpathians lowland into the northeastern steppe regions was not due to ecological niche differences but rather a result of other cultural factors. Furthermore, we highlighted that the populations of first Cucuteni-Trypillia groups had slightly more restricted ecological niches in the mid-Holocene ecosystems than Late Eneolithic groups. The results have significant implications to understand the geographical range and distribution of the last great Chalcolithic society of Old Europe and contribute to the characterization of ecological niches they have exploited during the cultural evolutionary process.