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Article ; Online: In Vivo

Rajawat, Yogendra S / Humbert, Olivier / Cook, Savannah M / Radtke, Stefan / Pande, Dnyanada / Enstrom, Mark / Wohlfahrt, Martin E / Kiem, Hans-Peter

Human gene therapy

2020 Volume 32, Issue 1-2, Page(s) 113–127

Abstract: Hematopoietic stem and progenitor cell (HSPC)- ... ...

Abstract	Hematopoietic stem and progenitor cell (HSPC)-based
MeSH term(s)	Animals ; Dogs ; Genetic Therapy ; Genetic Vectors/genetics ; Hematopoietic Stem Cells ; Humans ; Lentivirus/genetics ; Spumavirus ; Transduction, Genetic ; X-Linked Combined Immunodeficiency Diseases/genetics ; X-Linked Combined Immunodeficiency Diseases/therapy
Language	English
Publishing date	2020-09-23
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1028152-6
ISSN	1557-7422 ; 1043-0342
ISSN (online)	1557-7422
ISSN	1043-0342
DOI	10.1089/hum.2020.127
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Article: Effective Multi-lineage Engraftment in a Mouse Model of Fanconi Anemia Using Non-genotoxic Antibody-Based Conditioning.

Srikanthan, Meera A / Humbert, Olivier / Haworth, Kevin G / Ironside, Christina / Rajawat, Yogendra S / Blazar, Bruce R / Palchaudhuri, Rahul / Boitano, Anthony E / Cooke, Michael P / Scadden, David T / Kiem, Hans-Peter

Molecular therapy. Methods & clinical development

2020 Volume 17, Page(s) 455–464

Abstract: Conditioning chemotherapy is used to deplete hematopoietic stem cells in the recipient's marrow, facilitating donor cell engraftment. Although effective, a major issue with chemotherapy is the systemic genotoxicity that increases the risk for secondary ... ...

Abstract	Conditioning chemotherapy is used to deplete hematopoietic stem cells in the recipient's marrow, facilitating donor cell engraftment. Although effective, a major issue with chemotherapy is the systemic genotoxicity that increases the risk for secondary malignancies. Antibody conjugates targeting hematopoietic cells are an emerging non-genotoxic method of opening the marrow niche and promoting engraftment of transplanted cells while maintaining intact marrow cellularity. Specifically, this platform would be useful in diseases associated with DNA damage or cancer predisposition, such as dyskeratosis congenita, Schwachman-Diamond syndrome, and Fanconi anemia (FA). Our approach utilizes antibody-drug conjugates (ADC) as an alternative conditioning regimen in an FA mouse model of autologous transplantation. Antibodies targeting either CD45 or CD117 were conjugated to saporin (SAP), a ribosomal toxin.
Language	English
Publishing date	2020-02-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	2872938-9
ISSN	2329-0501 ; 2329-0501
ISSN (online)	2329-0501
ISSN	2329-0501
DOI	10.1016/j.omtm.2020.02.001
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Article: Autophagy in aging and in neurodegenerative disorders.

Rajawat, Yogendra S / Bossis, Ioannis

Hormones (Athens, Greece)

2008 Volume 7, Issue 1, Page(s) 46–61

Abstract: Autophagy (ATG) is the process of bulk degradation and recycling of long-lived proteins, macromolecular aggregates, and damaged intracellular organelles. Cellular homeostasis requires continuous removal of worn-out components and replacement with newly ... ...

Abstract	Autophagy (ATG) is the process of bulk degradation and recycling of long-lived proteins, macromolecular aggregates, and damaged intracellular organelles. Cellular homeostasis requires continuous removal of worn-out components and replacement with newly synthesized ones. Studies in yeast and other mammalian systems have increased our knowledge of the molecular mechanism of autophagy and the role of autophagy in various pathological conditions. Discovery of the genes involved in the process of autophagy has provided insight into the involvement of various molecular pathways. Growing evidence has indicated that diminished autophagic activity may play a pivotal role in the aging process. Cellular aging is characterized by a progressive accumulation of nonfunctional cellular components owing to oxidative damage and a decline in turnover rate and housekeeping mechanisms. Lysosomes are key organelles in the aging process due to their involvement in both macroautophagy and other housekeeping mechanisms. Autophagosomes themselves have limited degrading capacity and rely on fusion with lysosomes. Accumulation of defective mitochondria also appears to be critical in the progression of aging. Inefficient removal of nonfunctional mitochondria by lysosomes constitutes a major issue in the aging process. Autophagy has been associated with a growing number of pathological conditions, including cancer, myopathies, and neurodegenerative disorders. In this review, we discuss the cellular and molecular mechanisms involved in autophagy, the mechanisms of aging, and the possible role of autophagy in this process. Understanding the mechanisms by which autophagy impacts aging may provide useful molecular targets for pharmaceuticals designed to delay aging or correct conditions of premature aging.
MeSH term(s)	Aged ; Aging/physiology ; Autophagy/physiology ; Humans ; Lysosomes/physiology ; Neurodegenerative Diseases/pathology ; Neurons/pathology ; Neurons/physiology
Language	English
Publishing date	2008-03-10
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2075912-5
ISSN	1109-3099
ISSN	1109-3099
DOI	10.14310/horm.2002.1111037
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Article ; Online: Aging: central role for autophagy and the lysosomal degradative system.

Rajawat, Yogendra S / Hilioti, Zoe / Bossis, Ioannis

Ageing research reviews

2009 Volume 8, Issue 3, Page(s) 199–213

Abstract: The lysosomal network is the major intracellular proteolytic system accounting for more than 98% of long-lived bulk protein degradation and recycling particularly in tissues such as liver and muscles. Lysosomes are the final destination of intracellular ... ...

Abstract	The lysosomal network is the major intracellular proteolytic system accounting for more than 98% of long-lived bulk protein degradation and recycling particularly in tissues such as liver and muscles. Lysosomes are the final destination of intracellular damaged structures, identified and sequestered by the processes of macroautophagy and chaperone-mediated autophagy (CMA). In the process of macroautophagy, long-lived proteins and other macromolecular aggregates and damaged intracellular organelles are first engulfed by autophagosomes. Autophagosomes themselves have limited degrading capacity and rely on fusion with lysosomes. Unlike macroautophagy, CMA does not require intermediate vesicle formation and the cytosolic proteins recognized by this pathway are directly translocated to the lysosomal membrane. Aging is a universal phenomenon characterized by progressive deterioration of cells and organs due to accumulation of macromolecular and organelle damage. The continuous removal of worn-out components and replacement with newly synthesized ones ensures cellular homeostasis and delays the aging process. Growing evidence indicate that the rate of autophagosome formation and maturation and the efficiency of autophagosome/lysosome fusion decline with age. In addition, a progressive increase in intralysosomal concentration of free radicals and the age pigment lipofuscin further diminish the efficiency of lysosomal protein degradation. Therefore, integrity of the autophagosomal-lysosomal network appears to be critical in the progression of aging. Discovery of the genes involved in the process of autophagy has provided insight into the various molecular pathways that may be involved in aging and senescence. In this review, we discuss the cellular and molecular mechanisms involved in autophagy and the role of autophagosome/lysosome network in the aging process.
MeSH term(s)	Aging/metabolism ; Animals ; Autophagy/physiology ; Cellular Senescence/physiology ; Free Radicals/metabolism ; Humans ; Lipofuscin/metabolism ; Lysosomes/metabolism ; Oxidative Stress/physiology ; Proteins/metabolism ; Signal Transduction/physiology
Chemical Substances	Free Radicals ; Lipofuscin ; Proteins
Language	English
Publishing date	2009-07
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2075672-0
ISSN	1872-9649 ; 1568-1637
ISSN (online)	1872-9649
ISSN	1568-1637
DOI	10.1016/j.arr.2009.05.001
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Article ; Online: Rapid immune reconstitution of SCID-X1 canines after G-CSF/AMD3100 mobilization and in vivo gene therapy.

Humbert, Olivier / Chan, Frieda / Rajawat, Yogendra S / Torgerson, Troy R / Burtner, Christopher R / Hubbard, Nicholas W / Humphrys, Daniel / Norgaard, Zachary K / O'Donnell, Patricia / Adair, Jennifer E / Trobridge, Grant D / Scharenberg, Andrew M / Felsburg, Peter J / Rawlings, David J / Kiem, Hans-Peter

Blood advances

2018 Volume 2, Issue 9, Page(s) 987–999

Abstract: Hematopoietic stem-cell gene therapy is a promising treatment of X-linked severe combined immunodeficiency disease (SCID-X1), but currently, it requires recipient conditioning, extensive cell manipulation, and sophisticated facilities. With these ... ...

Abstract	Hematopoietic stem-cell gene therapy is a promising treatment of X-linked severe combined immunodeficiency disease (SCID-X1), but currently, it requires recipient conditioning, extensive cell manipulation, and sophisticated facilities. With these limitations in mind, we explored a simpler therapeutic approach to SCID-X1 treatment by direct IV administration of foamy virus (FV) vectors in the canine model. FV vectors were used because they have a favorable integration site profile and are resistant to serum inactivation. Here, we show improved efficacy of our in vivo gene therapy platform by mobilization with granulocyte colony-stimulating factor (G-CSF) and AMD3100 before injection of an optimized FV vector incorporating the human phosphoglycerate kinase enhancerless promoter. G-CSF/AMD3100 mobilization before FV vector delivery accelerated kinetics of CD3
MeSH term(s)	Animals ; Benzylamines ; CD4-CD8 Ratio ; Cyclams ; Disease Models, Animal ; Dog Diseases/blood ; Dog Diseases/genetics ; Dog Diseases/therapy ; Dogs ; Genetic Therapy ; Genetic Vectors/pharmacology ; Granulocyte Colony-Stimulating Factor/pharmacology ; Hematopoietic Stem Cell Mobilization ; Heterocyclic Compounds/pharmacology ; Humans ; Phosphoglycerate Kinase/genetics ; Spumavirus ; X-Linked Combined Immunodeficiency Diseases/blood ; X-Linked Combined Immunodeficiency Diseases/genetics ; X-Linked Combined Immunodeficiency Diseases/therapy ; X-Linked Combined Immunodeficiency Diseases/veterinary
Chemical Substances	Benzylamines ; Cyclams ; Heterocyclic Compounds ; Granulocyte Colony-Stimulating Factor (143011-72-7) ; Phosphoglycerate Kinase (EC 2.7.2.3) ; plerixafor (S915P5499N)
Language	English
Publishing date	2018-05-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2915908-8
ISSN	2473-9537 ; 2473-9529
ISSN (online)	2473-9537
ISSN	2473-9529
DOI	10.1182/bloodadvances.2018016451
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Article: HN protein of Newcastle disease virus causes apoptosis in chicken embryo fibroblast cells.

Ravindra, P V / Tiwari, Ashok K / Sharma, Bhaskar / Rajawat, Yogendra Singh / Ratta, Barkha / Palia, Sudesh / Sundaresan, N R / Chaturvedi, Uttara / Gangaplara, Arunakumar / Kumar, G B Aruna / Chindera, Kantaraja / Saxena, Meeta / Subudhi, P K / Rai, Anant / Chauhan, R S

Archives of virology

2008 Volume 153, Issue 4, Page(s) 749–754

Abstract: Newcastle disease virus (NDV), an avian paramyxovirus, induces apoptosis in chicken embryo fibroblast (CEF) cells. In the present investigation, the ability of haemagglutinin-neuraminidase (HN) protein of NDV to cause apoptosis in CEF cells was examined. ...

Abstract	Newcastle disease virus (NDV), an avian paramyxovirus, induces apoptosis in chicken embryo fibroblast (CEF) cells. In the present investigation, the ability of haemagglutinin-neuraminidase (HN) protein of NDV to cause apoptosis in CEF cells was examined. The results revealed that cells expressing the HN protein demonstrated decreased DNA content, phosphatidylserine exposure and increased cytoplasmic vacuolation. Up-regulation of caspase-1, -9, -8, -3, loss of mitochondrial transmembrane potential and an increase in oxidative stress were also observed in cells expressing the HN protein. Based on the above results it can be concluded that HN protein of NDV causes apoptosis in CEF cells.
MeSH term(s)	Animals ; Apoptosis ; Caspases/metabolism ; Cells, Cultured ; Chick Embryo/cytology ; Fibroblasts/metabolism ; Fibroblasts/pathology ; Fibroblasts/virology ; HN Protein/metabolism ; Newcastle disease virus/metabolism ; Newcastle disease virus/pathogenicity ; Oxidative Stress ; Phosphatidylserines/metabolism ; Up-Regulation
Chemical Substances	HN Protein ; Phosphatidylserines ; Caspases (EC 3.4.22.-)
Language	English
Publishing date	2008
Publishing country	Austria
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	7491-3
ISSN	1432-8798 ; 0304-8608
ISSN (online)	1432-8798
ISSN	0304-8608
DOI	10.1007/s00705-008-0057-2
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Article: HN protein of Newcastle disease virus causes apoptosis in chicken embryo fibroblast cells

Ravindra, P. V / Tiwari, Ashok K / Sharma, Bhaskar / Rajawat, Yogendra Singh / Ratta, Barkha / Palia, Sudesh / Sundaresan, N. R / Chaturvedi, Uttara / Aruna Kumar, G. B / Chindera, Kantaraja / Saxena, Meeta / Subudhi, P. K / Rai, Anant / Chauhan, R. S

Archives of virology. 2008 Apr., v. 153, no. 4

2008

Abstract	Newcastle disease virus (NDV), an avian paramyxovirus, induces apoptosis in chicken embryo fibroblast (CEF) cells. In the present investigation, the ability of haemagglutinin-neuraminidase (HN) protein of NDV to cause apoptosis in CEF cells was examined. The results revealed that cells expressing the HN protein demonstrated decreased DNA content, phosphatidylserine exposure and increased cytoplasmic vacuolation. Up-regulation of caspase-1, -9, -8, -3, loss of mitochondrial transmembrane potential and an increase in oxidative stress were also observed in cells expressing the HN protein. Based on the above results it can be concluded that HN protein of NDV causes apoptosis in CEF cells.
Keywords	DNA ; Newcastle disease virus ; Rubulavirus ; apoptosis ; caspase-1 ; chickens ; membrane potential ; oxidative stress
Language	English
Dates of publication	2008-04
Size	p. 749-754.
Publisher	Springer Vienna
Publishing place	Vienna
Document type	Article
ZDB-ID	7491-3
ISSN	1432-8798 ; 0304-8608
ISSN (online)	1432-8798
ISSN	0304-8608
DOI	10.1007/s00705-008-0057-2
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Article ; Online: Generation of a tumor vaccine candidate based on conjugation of a MUC1 peptide to polyionic papillomavirus virus-like particles.

Pejawar-Gaddy, Sharmila / Rajawat, Yogendra / Hilioti, Zoe / Xue, Jia / Gaddy, Daniel F / Finn, Olivera J / Viscidi, Raphael P / Bossis, Ioannis

Cancer immunology, immunotherapy : CII

2010 Volume 59, Issue 11, Page(s) 1685–1696

Abstract: Virus-like particles (VLPs) are promising vaccine technology due to their safety and ability to elicit strong immune responses. Chimeric VLPs can extend this technology to low immunogenicity foreign antigens. However, insertion of foreign epitopes into ... ...

Abstract	Virus-like particles (VLPs) are promising vaccine technology due to their safety and ability to elicit strong immune responses. Chimeric VLPs can extend this technology to low immunogenicity foreign antigens. However, insertion of foreign epitopes into the sequence of self-assembling proteins can have unpredictable effects on the assembly process. We aimed to generate chimeric bovine papillomavirus (BPV) VLPs displaying a repetitive array of polyanionic docking sites on their surface. These VLPs can serve as platform for covalent coupling of polycationic fusion proteins. We generated baculoviruses expressing chimeric BPV L1 protein with insertion of a polyglutamic-cysteine residue in the BC, DE, HI loops and the H4 helix. Expression in insect cells yielded assembled VLPs only from insertion in HI loop. Insertion in DE loop and H4 helix resulted in partially formed VLPs and capsomeres, respectively. The polyanionic sites on the surface of VLPs and capsomeres were decorated with a polycationic MUC1 peptide containing a polyarginine-cysteine residue fused to 20 amino acids of the MUC1 tandem repeat through electrostatic interactions and redox-induced disulfide bond formation. MUC1-conjugated fully assembled VLPs induced robust activation of bone marrow-derived dendritic cells, which could then present MUC1 antigen to MUC1-specific T cell hybridomas and primary naïve MUC1-specific T cells obtained from a MUC1-specific TCR transgenic mice. Immunization of human MUC1 transgenic mice, where MUC1 is a self-antigen, with the VLP vaccine induced MUC1-specific CTL, delayed the growth of MUC1 transplanted tumors and elicited complete tumor rejection in some animals.
MeSH term(s)	Animals ; Cancer Vaccines/therapeutic use ; Capsid Proteins/genetics ; Capsid Proteins/immunology ; Cattle ; Enzyme-Linked Immunosorbent Assay ; Humans ; Lymphocyte Activation ; Lymphoma, T-Cell/genetics ; Lymphoma, T-Cell/immunology ; Lymphoma, T-Cell/therapy ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Mucin-1/immunology ; Papillomaviridae/genetics ; Papillomavirus Vaccines/therapeutic use ; Peptide Fragments/genetics ; Peptide Fragments/immunology ; Peptide Fragments/metabolism ; Peptides/immunology ; T-Lymphocytes/immunology ; T-Lymphocytes, Cytotoxic/immunology ; Virion/genetics ; Virion/immunology
Chemical Substances	Cancer Vaccines ; Capsid Proteins ; L1 protein, Bovine papillomavirus ; MUC1 protein, human ; Mucin-1 ; Papillomavirus Vaccines ; Peptide Fragments ; Peptides ; polyarginine (25212-18-4)
Language	English
Publishing date	2010-07-21
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	195342-4
ISSN	1432-0851 ; 0340-7004
ISSN (online)	1432-0851
ISSN	0340-7004
DOI	10.1007/s00262-010-0895-0
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Article: Generation of a tumor vaccine candidate based on conjugation of a MUC1 peptide to polyionic papillomavirus virus-like particles

Pejawar-Gaddy, Sharmila / Rajawat, Yogendra / Hilioti, Zoe / Xue, Jia / Gaddy, Daniel F / Finn, Olivera J / Viscidi, Raphael P / Bossis, Ioannis

Cancer immunology, immunotherapy. 2010 Nov., v. 59, no. 11

2010

Abstract	Virus-like particles (VLPs) are promising vaccine technology due to their safety and ability to elicit strong immune responses. Chimeric VLPs can extend this technology to low immunogenicity foreign antigens. However, insertion of foreign epitopes into the sequence of self-assembling proteins can have unpredictable effects on the assembly process. We aimed to generate chimeric bovine papillomavirus (BPV) VLPs displaying a repetitive array of polyanionic docking sites on their surface. These VLPs can serve as platform for covalent coupling of polycationic fusion proteins. We generated baculoviruses expressing chimeric BPV L1 protein with insertion of a polyglutamic-cysteine residue in the BC, DE, HI loops and the H4 helix. Expression in insect cells yielded assembled VLPs only from insertion in HI loop. Insertion in DE loop and H4 helix resulted in partially formed VLPs and capsomeres, respectively. The polyanionic sites on the surface of VLPs and capsomeres were decorated with a polycationic MUC1 peptide containing a polyarginine-cysteine residue fused to 20 amino acids of the MUC1 tandem repeat through electrostatic interactions and redox-induced disulfide bond formation. MUC1-conjugated fully assembled VLPs induced robust activation of bone marrow-derived dendritic cells, which could then present MUC1 antigen to MUC1-specific T cell hybridomas and primary naïve MUC1-specific T cells obtained from a MUC1-specific TCR transgenic mice. Immunization of human MUC1 transgenic mice, where MUC1 is a self-antigen, with the VLP vaccine induced MUC1-specific CTL, delayed the growth of MUC1 transplanted tumors and elicited complete tumor rejection in some animals.
Keywords	immunotherapy
Language	English
Dates of publication	2010-11
Size	p. 1685-1696.
Publisher	Springer-Verlag
Publishing place	Berlin/Heidelberg
Document type	Article
ZDB-ID	195342-4
ISSN	1432-0851 ; 0340-7004
ISSN (online)	1432-0851
ISSN	0340-7004
DOI	10.1007/s00262-010-0895-0
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