Ht, The NetherlandsAbstractCancer is usually viewed as a caricature of typical developmental processes, however the extent by which its cellular heterogeneity really recapitulates multi-lineage differentiation processes of normal tissues remains unknown. Here, we implement “single-cell PCR gene-expression analysis” (SINCE-PCR) to dissect the cellular NFPS Membrane Transporter/Ion Channel composition of principal human typical colon and colon cancer epithelia. We show that human colon cancer tissues contain distinct cell populations whose transcriptional identities mirror those of your various cellular lineages of regular colon. By generating monoclonal tumor xenografts from injection of a single-cell (n = 1), we show that transcriptional diversity of cancer tissues is largely explained by in vivo multi-lineage differentiation, not just by clonal genetic heterogeneity. Ultimately, we show that perturbations in gene-expression programs linked to multi-lineage differentiation strongly associate with patient survival. Guided by SINCE-PCR data, we develop two-gene classifier systems (KRT20 vs CA1, MS4A12, CD177, SLC26A3) that predict clinical outcomes with hazard-ratios superior to pathological grade and comparable to microarray-derived multi-gene expression signatures.Users may possibly view, print, copy, download and text and data- mine the content material in such documents, for the purposes of academic investigation, topic generally for the full Conditions of use: http://nature.com/authors/editorial_policies/license.html#terms Correspondence to: Hexestrol Autophagy Stephen R. Quake, Ph.D., Professor of Bioengineering and Applied Physics, Stanford University, Clark Center, E350Q, 318 Campus Drive, Stanford, California, 94305, phone (650) 736-7890, fax(650) 736-1961, [email protected]. These authors contributed equally to the study. AUTHOR CONTRIBUTIONS P.D., T.K., D.S., M.F.C. and S.R.Q. conceived the study and created the experiments. P.S.R., M.E.R., A.A.L., M.Z., N.F.N, M. v. d. W. and H.C. offered intellectual guidance within the style of selected experiments. P.D., T.K., D.S., P.S.R., A.A.L., S.S., J.O., D.M.J., D.Q., J.W., Y.S. and S.H. performed the experiments. P.D., T.K., D.S., N.F.N., Y.S., M.F.C. and S.R.Q analyzed the data and/or supplied intellectual guidance in their interpretation. J.B., A.A.S. and B.V. offered samples and reagents. P.D., T.K., D.S., M.F.C. and S.R.Q. wrote the paper.Dalerba et al.PageThe in vivo cellular composition of solid tissues is usually difficult to investigate within a comprehensive and quantitative way. Approaches for instance immunohistochemistry and flow cytometry are limited by the availability of antigen-specific monoclonal antibodies and by the tiny quantity of parallel measurements that can be performed on each and every individual cell. Classic high-throughput assays, which include gene-expression arrays, when performed on entire tissues, deliver info on average gene expression levels, and can be only indirectly correlated to quantitative modifications in cellular subpopulations. These limitations become especially difficult to overcome when studying minority populations, including stem cells, whose identification is produced elusive by their low numbers and by the lack of exclusive markers. Furthermore, in pathological states, for instance cancer, it is frequently impossible to decide no matter whether perturbations in gene expression detected in whole tissues are due to modifications within the relative composition of diverse cell kinds or to aberrations within the gene-expression profile of mutated cells. By way of example, while.