Transplantation of endothelial cells (ECs) is a promising therapeutic approach for ischemic disorders. In addition, the generation of ECs has become increasingly important for providing vascular plexus to regenerated organs, such as the liver. Although many attempts have been made to generate ECs from pluripotent stem cells and nonvascular cells, the minimum number of transcription factors that specialize in directly inducing vascular ECs remains undefined. Here, by screening 18 transcription factors that are important for both endothelial and hematopoietic development, we demonstrate that ets variant 2 (ETV2) alone directly converts primary human adult skin fibroblasts into functional vascular endothelial cells (ETVECs). In coordination with endogenous FOXC2 in fibroblasts, transduced ETV2 elicits expression of multiple key endothelial development factors, including FLI1, ERG, and TAL1, and induces expression of endothelial functional molecules, including EGFL7 and von Willebrand factor. Consequently, ETVECs exhibits EC characteristics in vitro and forms mature functional vasculature in Matrigel plugs transplanted in NOD SCID mice. Furthermore, ETVECs significantly improve blood flow recovery in a hind limb ischemic model using BALB/c-nu mice. Our study indicates that the creation of ETVECs provides further understanding of human EC development induced by ETV2.uman vascular endothelial cells (ECs) generated from pluripotent stem cells (PSCs), including embryonic stem cells and induced PSCs (iPSCs), or nonvascular cells have great therapeutic potential for treating ischemic vascular diseases (1, 2). In addition, the generation of ECs has become increasingly important for providing vascular plexus to regenerated organs, such as the liver (3). iPSCs are an especially promising source for creating ECs, although the main limitation of applying iPSCs is the risk of incomplete differentiation and tumorigenicity (4, 5). To bypass the fully pluripotent state, multiple research groups have generated ECs by culturing fibroblasts transduced with iPSCinducing factors (OCT4, SOX2, KLF4, and c-MYC) under defined EC culture conditions (6, 7). The long-term stability of these ECs remains to be established, however; the PSC-derived ECs often show poor proliferative abilities and drift into nonvascular lineages (8).Combined expression of multiple transcription factors specific to a particular lineage has been demonstrated to change somatic cell fate by bypassing pluripotency; for example, Gata4, Mef2c, and Tbx5 convert murine cardiac fibroblasts into functional cardiomyocytes (9), and Ascl1, Brn2, and Myt1 induce neurons from murine fibroblasts (10). This "direct lineage conversion" approach offers promising prospects for creating cells of biomedical interest for cellular replacement therapies. This approach is also useful for studying the physiological mechanisms of transcriptional reprogramming, such as the establishment of cellular identity, and the transcriptional regulatory networks that drive terminal differentiation and fu...

A431 cells have an amplification of the epidermal growth factor (EGF) receptor gene, the cellular homolog of the v-erb B oncogene, and overproduce an aberrant 2.9-kilobase RNA that encodes a portion of the EGF receptor. A cDNA (pE15) for the aberrant RNA was cloned, sequenced, and used to analyze genomic DNA blots from A431 and normal cells. These data indicate that the aberrant RNA is created by a gene rearrangement within chromosome 7, resulting in a fusion of the 5' portion of the EGF receptor gene to an unidentified region of genomic DNA. The unidentified sequences are amplified to about the same degree (20-to 30.fold) as the EGF receptor sequences. In situ hybridization to chromosomes from normal cells and A431 cells show that both the EGF receptor gene and the unidentified DNA are localized to the pl4-pi2 region of chromosome 7. By using cDNA fragments to probe DNA blots from mouse-A431 somatic cell hybrids, the rearranged receptor gene was shown to be associated with translocation chromosome M4.

We previously reported that the structural gene for epidermal growth factor receptor (EGFR) can be mapped to the p22→qter region of human chromosome 7 (Shimizu et al., 1979, 1980). In the present study, we produced two series of human-mouse cell hybrids by fusing mouse A9 cells that are deficient in EGFR with the human diploid fibroblast lines GM1356, 46, XX, t(l;7)(p34;pl3), and GM2068, 46, XX, t(6;7)(q27;q22), both of which possess EGF receptors. Expression of EGF binding ability in the former series of cell hybrids was correlated with the retention of the human translocation chromosome containing the 7p13→qter region, and in the latter series of cell hybrid it was correlated with the retention of the human translocation chromosome containing the 7pter→q22 region. Therefore, the EGFR gene can be localized in the p13→q22 region of chromosome 7.

Psoriasis is considered a Th17-type autoimmune skin inflammatory disease; however, involvement of an autoantigen-specific TCR has not been established. In this study, we show that psoriasis-like skin inflammation can be induced by autoreactive Th17 cells. We previously developed the desmoglein 3–specific TCR-transgenic (Dsg3H1) mouse, in which CD4+ T cells recognize physiological epidermal autoantigen. T cells from Dsg3H1 mice were polarized into Th17 cells in vitro and then adoptively transferred into Rag2−/− mice. Dsg3H1-Th17 cells induced severe psoriasis-like skin inflammation within 2 wk after transfer in the tissues in which desmoglein 3 is expressed. Such pathology was not observed when wild-type Th17 cells or Th1-skewed Dsg3H1 T cells were transferred, and it was strongly suppressed by anti–IL-12/23 and anti–IL-17 Abs. Although IFN-γ+/IL-17+ T cells accumulated in the skin lesions of mice that received Dsg3H1-Th17 cells, IFN-γ–deficient Dsg3H1-Th17 cells were fully pathogenic. These results demonstrate that cutaneous psoriasis-like immunopathology can be developed by epidermis-specific recognition of Th17 cells, which is strictly dependent on IL-17 but not IFN-γ.

Calcium dependent proteases (calpains, CAPNs, E.C.3.4.22.17) constitute a family of proteins which share a homologous cysteine-protease domain (large subunits, L1 L2, and L3) and an E-F hand Ca²⁺-binding domain (L1, L2, L3, and small subunit, S). We have mapped the genes for four calpain proteins (L1 L2, L3, and S) on four distinct human chromosomes by a combination of spot-blot hybridization to flow-sorted chromosomes and Southern hybridization of DNAs from a human × mouse hybrid cell panel. The genes for calpain L1 (CAPNl, large subunit of calpain I), L2 (CAPN2, large subunit of calpain II), L3 (CAPN3, a protein related to the large subunits), and S (CAPN4, a small subunit common to calpains I and II) were assigned to human chromosomes 11, 1, 15, and 19, respectively.

SP-40,40 is a serum glycoprotein consisting of two different subunits (α and β) assembled into a dimer by disulfide bonds. Northern blot hybridization, using total RNA from several cell lines, showed that SP-40,40 is expressed in glioblastoma and testicular tumor cells, as well as hepatoma cells. Spot blot hybridization of flow-sorted human chromosomes, using a SP-40,40 cDNA fragment as a probe, localized the gene for SP-40,40 to human chromosome 8. This gene has been given the designation CLI, for complement lysis inhibitor, by the Human Gene Nomenclature Committee.

Fluorescent in situ hybridization in combination with Q-banding revealed that the human xanthine dehydrogenase (XDH) gene is located on band p23 of chromosome 2.

Inter-α-trypsin inhibitor family heavy chain-related protein (IHRP) is a novel glycoprotein isolated from human plasma. The cDNA encoding IHRP has recently been cloned from human liver cDNA libraries. We report the mapping of this gene (ITIHL1) by fluorescence in situ hybridization using a 2.5-kb cDNA fragment as a probe. ITIHL1 was localized to chromosome region 3p21→p 14 where the genes of heavy chain 1 and 3 of inter-α-trypsin inhibitor are located. This result, together with significant homology between the nucleotide sequences of ITIHL1 and the heavy chain genes, supports ITIHL1 as being a member of an evolutionary related gene family of ITI heavy chains. Northern blot analysis indicated that IHRP was predominantly synthesized in liver. From Southern blot analysis, it was tentatively concluded that ITIHL1 is a single copy gene.