Deaths from cancer, which affect more than eight million people worldwide every year, are largely due to metastasis, the spread of a cancer from its native site to other organs. A vital part of the metastatic cascade is the initial dissemination of cancer cells from a confined lesion into neighboring tissue through cell migration and invasion. In addition to biochemical cues, the role of physical cues in the microenvironment in directing migration is particularly relevant for certain solid tumors with a characteristic fibroinflammatory response called desmoplasia. Desmoplasia, such as those in breast and pancreatic ductal carcinomas, are often associated with increased deposition of collagen-I, a fibrous protein of the extracellular matrix (ECM) that not only confers increased stiffness to the tumor mass but also provides a fiber architecture recognizable by cellular sensory complexes. Specifically, live imaging of breast tumors and archival tissue imaging have established that aligned collagen tracks provide contact guidance cues for directed cancer cell migration and dissemination in mature breast tumors, leading to increased invasion, metastasis and decreased disease-free survival in patients. Using multiphoton imaging of archival and live tumor samples from mouse models and human patients, we explored the spatio-temporal evolution of such architectures in pancreatic ductal adenocarcinoma (PDA), another highly desmoplastic disease. Our findings demonstrate that organized tumor-associated collagen architectures develop in the PDA stroma even at the pre-invasive stage and both undifferentiated single cells and cancer cells within well differentiated epithelial clusters interact with these aligned collagen tracks in vivo. Mimicking the aligned collagen networks with micropatterned substrates we studied contact guidance in a panel of phenotypically diverse breast and pancreatic carcinoma cells. We demonstrate that for single cells, aligned architectures induce constrained focal adhesion maturation and associated F-actin alignment, consequently orchestrating anisotropic traction stresses that drive cell orientation and directional migration. While such interactions allow single mesenchymal-like cells to spontaneously “sense” and follow topographic alignment, intercellular interactions within epithelial clusters counteract anisotropic cell-substratum forces, resulting in substantially lower directional response. Further analyses indicate that anisotropic cell-ECM interactions from organized periductal collagen may indeed contribute to cell extrusion from ductal epithelia into the stroma leading to enhanced and early dissemination in pancreatic cancer. According to our findings, such contact guided spreading of cancer cells may be inhibited most efficiently by dismantling the fiber architecture or diminishing its density, as well as by reducing the strength of cell-ECM interactions. To validate such findings and further establish causal relationships, we engineered novel in vitro 3D substrates using a simple method to align collagen gels by guided cellular compaction. Our method yielded highly aligned, acellular collagen constructs as a controlled microenvironment for in vitro experiments. Further, we integrated such aligned collagen matrices to cell dense tumor-like plugs, allowing tracking of the temporal evolution of the advancing invasion fronts over several days. Live cell imaging and analysis of 3D cell migration revealed profoundly enhanced motility in aligned collagen matrices for the putative subpopulation of carcinoma cells with high metastatic capacity, termed cancer stem cells. Heterogeneity in cell migration behavior was also observed between cells at the leading edge and those within the tumor boundary, thus demonstrating the versatility of these platforms in capturing the dynamics of contact guided carcinoma dissemination. Overall, our findings elucidate how phenotypically diverse cancer cells perceive ECM alignment at the molecular level. Such anisotropic interactions likely contribute to single cell extrusion and dissemination in early pancreatic tumors and our findings provide a framework to design therapeutic targeting of the same. Further, we also develop 3D in vitro systems which are ideal to dissect, validate and refine our current understanding of this critical pathway to metastasis.