Granulocyte Colony-Stimulating Factor Expression from Transduced Vascular Smooth Muscle Cells Provides Sustained Neutrophil Increases in Rats

Lejnieks, Daniel V.; Han, Sang Won; Ramakrishnan, Nagarajan; Lau, Stella; Osborne, William

doi:10.1089/hum.1996.7.12-1431

Cited by 17 publications

(12 citation statements)

References 30 publications

(32 reference statements)

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“…[57][58][59] Vascular smooth muscle cells, to engraft in humans, may necessitate arterial injury. [60][61][62] Hematopoietic stem cells can be difficult to expand in culture and gene-modify, 63,64 and very large numbers are required for engraftment in the absence of a toxic 'conditioning' regimen. 65 Lymphocytes possess a short lifespan, 66 and human umbilical vein endothelial cells cannot be obtained from an adult.…”

Section: Discussionmentioning

confidence: 99%

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

et al. 2003

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Marrow stromal cells (MSCs) are postnatal progenitor cells that can be easily cultured ex vivo to large amounts. This feature is attractive for cell therapy applications where genetically engineered MSCs could serve as an autologous cellular vehicle for the delivery of therapeutic proteins. The usefulness of MSCs in transgenic cell therapy will rely upon their potential to engraft in nonmyeloablated, immunocompetent recipients. Further, the ability to deliver MSCs subcutaneously -as opposed to intravenous or intraperitoneal infusions -would enhance safety by providing an easily accessible, and retrievable, artificial subcutaneous implant in a clinical setting. To test this hypothesis, MSCs were retrovirally engineered to secrete mouse erythropoietin (Epo) and their effect was ascertained in nonmyeloablated syngeneic mice. Epo-secreting MSCs when administered as 'free' cells by subcutaneous or intraperitoneal injection, at the same cell dose, led to a significant -yet temporaryhematocrit increase to over 70% for 55713 days. In contrast, in mice implanted subcutaneously with Matrigeltembedded MSCs, the hematocrit persisted at levels 480% for over 110 days in four of six mice (Po0.05 logrank). Moreover, Epo-secreting MSCs mixed in Matrigel elicited and directly participated in blood vessel formation de novo reflecting their mesenchymal plasticity. MSCs embedded in human-compatible bovine collagen matrix also led to a hematocrit 470% for 7578.9 days. In conclusion, matrixembedded MSCs will spontaneously form a neovascularized organoid that supports the release of a soluble plasma protein directly into the bloodstream for a sustained pharmacological effect in nonmyeloablated recipients.

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Section: Discussionmentioning

confidence: 99%

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

et al. 2003

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“…43 Recombinant canine G-CSF (Amgen, Thousand Oaks, CA) was used to construct a proliferation-response curve with murine NFS-60 cells. 43,44 Proliferation was assayed by a commercial dye production assay (Cell Titer 96; Promega, Madison, WI). In brief, NFS-60 cells seeded at a concentration of 10 5 cells/well in 96-well microtiter plates were cultured for 24 hours at 37°C, 5% CO 2 .…”

mentioning

confidence: 99%

Treatment of canine cyclic neutropenia by lentivirus-mediated G-CSF delivery

Yanay

Barry

Katen

et al. 2003

Blood

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Cyclic neutropenia is a rare disease that occurs both in humans and gray collie dogs and is characterized by recurrent severe neutropenia leading to bacterial infections and shortened life expectancy. Daily injections of recombinant granulocyte colony-stimulating factor (rG-CSF) are effective in shortening the period of severe neutropenia and reducing infections. After demonstrating that rG-CSF induced elevated neutrophil production in an affected dog, cytokine administration was stopped and 10 9 infectious units (IUs) of a lentivirus pseudotyped with vesicular stomatitis virus G protein (VSV-G) encoding canine G-CSF cDNA was administered intramuscularly. Serial blood cell counts showed elevated neutrophil production for longer than 17 months. Although neutrophil counts continued to cycle, the range at nadirs was from 3710 to 5300 cells/L, well above the nadirs before lentivirus administration. After the injection of lentivirus, mean neutrophil counts ؎ SD were 12 460 ؎ 4240 cells/ L, significantly increased over both pretreatment values of 3040 ؎ 2540 cells/L (P < .0001) and neutrophil counts during G-CSF administration of 10 290 ؎ 4860 cells/L (P < .007). The changes in blood counts from lentivirus injection were associated with absence of clinical signs of infection and fever. The gray collie continued to gain weight and was no longer housed in a pathogen-free environment. Genomic DNA from muscle at injection sites was positive for provirus, whereas gonad, lung, spleen, heart, liver, kidney, leukocytes, and noninjected muscle samples were all negative for provirus. Thus, intramuscular administration of lentivirus encoding G-CSF provided sustained therapeutic levels of neutrophils, suggesting this approach may be applied for long-term treatment of patients with cyclic and other neutropenias. IntroductionCyclic neutropenia is a rare disease that occurs both in man and gray collie dogs. In collie dogs, cyclic hematopoiesis is inherited as an autosomal recessive disease. 1,2 The dogs have 12-to 14-day cyclic variations in blood neutrophils, monocytes, lymphocytes, eosinophils, reticulocytes, and platelets due to periodic fluctuations in blood cell production by the bone marrow. 1,2 The recurrent severe neutropenia leads to bacterial infections and shortened life expectancy. The disorder can be cured by bone marrow transplantation from a normal dog to a gray collie as well as transferred from a gray collie to a normal littermate. 3,4 This transplantability strongly supports the concept that this is a disease of defective regulation of hematopoietic stem cells.Gray collie dogs with cyclic hematopoiesis have been successfully treated with granulocyte colony-stimulating factor (G-CSF). 5,6 The severe recurrent neutropenia in gray collie dogs was not abrogated by in vivo interleukin 3 (IL-3) or granulocytemacrophage colony-stimulating factor (GM-CSF) treatment. 5 In contrast, G-CSF prevented the recurrent neutropenia and obliterated periodic fluctuation of monocyte, eosinophil, reticulocyte, and platelet counts. 5,6 ...

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“…One microliter of pRRL-cPPT-CMV-cGCSF-PRE-SIN virus (6 · 10 8 IU/ml) was added to 1 ml of test serum, incubated at room temperature for 1 hr, filtered through a 0.2-lm (pore size) syringe filter, and added with DEAE-dextran, at a final concentration of 10 lg/ml, to a 100-mm dish containing 10 4 293T cells. The next day the medium was changed and 3 days later the medium was collected and passed through a 0.2-lm (pore size) syringe filter, and the G-CSF concentration was measured in a murine NFS-60 cell proliferation assay (Lejnieks et al, 1996;Yanay et al, 2003). Recombinant canine G-CSF (rcG-CSF) was used as a positive control.…”

Section: Virus-neutralizing Assaymentioning

confidence: 99%

“…Anticoagulated blood samples (0.5 ml) were obtained serially from a peripheral vein and total white blood cell (WBC), platelet, and hematocrit values were determined with an automated blood cell counter (Coulter, Hialeah, FL). In some cases differential WBC counts were performed manually (Lejnieks et al, 1996).…”

Section: Dog Care Virus Administration and Blood Drawingmentioning

confidence: 99%

Repeated Lentivirus-Mediated Granulocyte Colony-Stimulating Factor Administration to Treat Canine Cyclic Neutropenia

2012

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Granulocyte Colony-Stimulating Factor Expression from Transduced Vascular Smooth Muscle Cells Provides Sustained Neutrophil Increases in Rats

Cited by 17 publications

References 30 publications

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

Treatment of canine cyclic neutropenia by lentivirus-mediated G-CSF delivery

Repeated Lentivirus-Mediated Granulocyte Colony-Stimulating Factor Administration to Treat Canine Cyclic Neutropenia

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