Dna Vec Research Inc.

Dnavec Corporation

Other

Tanabe Seiyaku Company, Limited

Yasuji Ueda

Makoto Inoue

Mamoru Hasegawa

Akihiro Iida

Masanori Kobayashi

Yoshikazu Yonemitsu

Hiroaki Kinoh

Yasushi Kondo

Toshihiro Nakajima

Hiroshi Ban

Yui Harada

Masanori Miyazaki

Noemi Fusaki

Yutaka Hanazono

Makoto Asakawa

Katsuo Sueishi

Koichi Saeki

Tsugumine Shu

Yutaka Suzuki

Yasuhiro Ikeda

Toshiaki Tabata

Kenji Nakamaru

Tsuyoshi Tokusumi

Takahiro Hirata

Hiroyuki Sakakibara

Kaio Kitazato

Hidekazu Kuma

Fumino Tokito

Yumiko Tokusumi

Mamoru c/o Dnavec Corporation Hasegawa

Tsugumine c/o Dnavec Corporation Shu

Hiroto c/o Dnavec Corporation Hara

The present invention provides Sendai virus vectors in which genes that encode reprograming factors for inducing pluripotent stem cells are incorporated in a specific order, compositions comprising these vectors for gene delivery to be used in the induction of pluripotent stem cells, and uses thereof. Incorporation of the KLF gene, OCT gene, and SOX gene in a specific order into a single Sendai virus vector successfully and significantly increased the efficiency of pluripotent stem cell induction. Loading multiple reprogramming factors into a single vector can further increase the induction efficiency of pluripotent stem cells while reducing the number of necessary vectors.

Composition for inducing pluripotent stem cell, and use thereof

An objective of the present invention is to provide methods for producing dendritic cells (DCs), which comprise the step of culturing DC precursor cells in the presence of a plurality of cytokines, produced dendritic cells, and uses thereof. The present inventors discovered that dendritic cells with a high IL-12 productivity can be obtained by culturing DC precursor cells in the presence of a plurality of cytokines, followed by about one week of culture in the presence of GM-CSF and IL-4. The present invention enables preparation of a large amount of DCs with a high IL-12 productivity from a small number of DC precursor cells, and therefore makes it easier to increase the number of DCs for administration in DC-based anti-tumor immune therapy, treatment of infections, etc. Thus, the effect of DC vaccines is expected to be enhanced.

Method for producing dendritic cells

The present invention provides novel methods for treating diseases associated with apoptotic degeneration in ocular tissue cells by effective administration of pigment epithelium derived factor (PEDF). The present inventors studied PEDF as a means to prevent ganglion cell death, the final pathology of glaucoma. The present invention is particularly focused on SIV vectors for effective methods for delivering PEDF, and constructed an SIV-PEDF vector. When the SIV-PEDF vector was administered subretinally to an ischemia reperfusion model and NMDA-induced model, a significant suppression effect on ganglion cell death was observed. The present inventors therefore discovered that the SIV-PEDF vector is an effective pharmaceutical agent for treating diseases associated with apoptotic degeneration in ocular tissue cells, such as glaucoma.

Therapeutic agents for diseases associated with apoptotic degeneration in ocular tissue cells that use SIV-PEDF vectors

The present invention provides cell fusogenic vectors having replicative ability, whose protease-dependent tropism has been modified. M gene-deficient viral vectors encoding modified F proteins, in which the cleavage site of the F protein of paramyxovirus is modified to be cleaved by different proteases, were produced. In cells transfected with these vectors, the genomic RNA present in the vectors is replicated, and cell fusogenic infection spreads to neighboring cells depending on the presence of other proteases; however, no viral particles are released. The vectors of this invention, encoding the F proteins which are cleaved by proteases whose activity is enhanced in cancer, show cancer growth suppressive effect in vivo.

Vectors with modified protease-dependent tropism

The present invention provides a retroviral vector containing a membrane protein having a hemagglutinin activity. The present inventors constructed a retroviral vector pseudotyped by the membrane protein having a hemagglutinin activity. This viral vector showed gene transfer at a high efficiency into host cells. In particular, it was established that genes can be transferred thereby at a high efficiency into cells into which genes can hardly be transferred by the conventional techniques, for example, blood cells and hematopoietic cells including hematopoietic stem cells, and mucous cells including mucosa epithelial cells. The viral vector of the present invention is highly useful as a vector for gene therapy.

Pseudotype retroviral vectors containing membrane proteins having hemagglutinin activity

The present invention provides cell fusogenic vectors having replicative ability, whose protease-dependent tropism has been modified. M gene-deficient viral vectors encoding modified F proteins, in which the cleavage site of the F protein ofis modified to be cleaved by different proteases, were produced. In cells transfected with these vectors, the genomic RNA present in the vectors is replicated, and cell fusogenic infection spreads to neighboring cells depending on the presence of other proteases; however, no viral particles are released. The vectors of this invention, encoding the F proteins which are cleaved by proteases whose activity is enhanced in cancer, show cancer growth suppressive effect in vivo.

Highly efficient gene transfer into primate-derived embryonic stem (ES) cells has successfully been achieved by using a simian immunodeficiency virus vector (SIV) pseudotyped with VSV-G protein, which is a surface glycoprotein of vesicular stomatitis virus (VSV) The present invention provides simian immunodeficiency virus vectors for gene transfer to primate ES cells. The method for gene transfer to primate ES cells using the vectors of the present invention is useful in, for example, research into embryology and disease, clinical applications, and experimental models for primates. The method is also useful in assaying and screening for genes and reagents able to enhance the specific differentiation of tissues or cells, and which are useful in preparing desired cells or tissues differentiated from ES cells.

Gene transfer into primate embryonic stem cells using VSV-G pseudotyped simian immunodeficiency virus vectors

F gene-deficient virus virions are successfully recovered by using an F gene-deficient Sendai virus genomic cDNA. Further, F gene-deficient infectious viral particles are successfully constructed by using F-expressing cells as helper cells. Also, F gene and HN gene-deficient virus virions are successfully recovered by using a virus genomic cDNA deficient in both F gene and HN gene. Further, F gene and HN gene-deficient infectious viral particles are successfully produced by using F- and HN-expressing cells as helper cells. A virus deficient in F gene and HN gene and having F protein is constructed by using F-expressing cells as helper cells. In addition, M gene-deficient infectious virus particles were produced using helper cells expressing M protein. From cells infected with M gene-deficient viruses, release of virus-like particles was inhibited. Further, a VSV-G pseudo type virus is successfully constructed by using VSV-G-expressing cells. Techniques for constructing these deficient viruses contribute to the development of vectors of Paramyxoviridae usable in gene therapy.

Envelope gene-deficient Paramyxovirus vector

Growing community of inventors

Ibaraki, Japan

Yasuji Ueda