Expression of Human Calcitonin by Microencapsulated Recombinant Myoblasts

Wang, Yeming; Zeng, Bingfang; Li, Xin

doi:10.1007/s10529-006-9115-6

Cited by 8 publications

(10 citation statements)

References 13 publications

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“…43 In another study, investigators compared the toxic effects of actinomycin-D encapsulated either in the aqueous phase or in the lipid phase of liposomes and the nonencapsulated actinomycin-D on the blood forming system on cell proliferation in the intestine and on antibody production by spleen lymphocytes and have confirmed that actinomycin-D is less toxic to mice than nonencapsulated actinomycin-D, but retains its tumoricidal activity. 44 Similarly, nerve growth factor has also been encapsulated into liposomes in order to protect it from the enzyme degradation in vivo and promote permeability across the blood-brain barrier. 45 Furthermore, MEND has been developed as a novel nonviral gene delivery system.…”

Section: Mts and Liposome-encapsulated Stem Cellsmentioning

confidence: 99%

Clinical significance of metallothioneins in cell therapy and nanomedicine

Sharma¹,

Rais²,

Sandhu³

et al. 2013

IJN

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Mammalian metallothioneins (MTs) are low molecular weight (6-7 kDa) cysteinerich proteins that are specifically induced by metal nanoparticles (NPs). MT induction in cell therapy may provide better protection by serving as antioxidant, anti-inflammatory, antiapoptotic agents, and by augmenting zinc-mediated transcriptional regulation of genes involved in cell proliferation and differentiation. Liposome-encapsulated MT-1 promoter has been used extensively to induce growth hormone or other genes in culture and gene-manipulated animals. MTs are induced as a defensive mechanism in chronic inflammatory conditions including neurodegenerative diseases, cardiovascular diseases, cancer, and infections, hence can serve as early and sensitive biomarkers of environmental safety and effectiveness of newly developed NPs for clinical applications. Microarray analysis has indicated that MTs are significantly induced in drug resistant cancers and during radiation treatment. Nutritional stress and environmental toxins (eg, kainic acid and domoic acid) induce MTs and aggregation of multilamellar electron-dense membrane stacks (Charnoly body) due to mitochondrial degeneration. MTs enhance mitochondrial bioenergetics of reduced nicotinamide adenine dinucleotide-ubiquinone oxidoreductase (complex-1), a rate-limiting enzyme complex involved in the oxidative phosphorylation. Monoamine oxidase-B inhibitors (eg, selegiline) inhibit α-synuclein nitration, implicated in Lewy body formation, and inhibit 1-methyl 4-phenylpyridinium and 3-morpholinosydnonimineinduced apoptosis in cultured human dopaminergic neurons and mesencephalic fetal stem cells. MTs as free radical scavengers inhibit Charnoly body formation and neurodegenerative α-synucleinopathies, hence Charnoly body formation and α-synuclein index may be used as early and sensitive biomarkers to assess NP effectiveness and toxicity to discover better drug delivery and surgical interventions. Furthermore, pharmacological interventions augmenting MTs may facilitate the theranostic potential of NP-labeled cells and other therapeutic agents. These unique characteristics of MTs might be helpful in the synthesis, characterization, and functionalization of emerging NPs for theranostic applications. This report highlights the clinical significance of MTs and their versatility as early, sensitive biomarkers in cell-based therapy and nanomedicine.

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Section: Mts and Liposome-encapsulated Stem Cellsmentioning

confidence: 99%

Clinical significance of metallothioneins in cell therapy and nanomedicine

Sharma¹,

Rais²,

Sandhu³

et al. 2013

IJN

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“…Gene therapy has emerged as one of the most rapidly advancing fields of biotechnology with great promise for treating various diseases (Jung et al 2005;Wang et al 2006). Bone regeneration needs to be enhanced when treating fracture non-unions and healing large bone defects due to tumor or trauma.…”

Section: Introductionmentioning

confidence: 99%

Osteogenic differentiation effects on rat bone marrow-derived mesenchymal stromal cells by lentivirus-mediated co-transfection of human BMP2 gene and VEGF165 gene

2007

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We proposed a novel combined gene therapy of human vascular endothelial growth factor 165 gene (hVEGF165) and human bone morphogenetic protein 2 gene (hBMP2) for bone regeneration by lentivirus-mediated co-transfection of both genes into rat bone marrow-derived mesenchymal stromal cells (MSCs). Both genes were successfully co-expressed in MSCs confirmed by real-time PCR and ELISA. And the alkaline phosphatase activity of MSCs was significantly augmented by the co-transfection with both genes than any single gene transfection (P < 0.01). These results demonstrated the feasibility of the combined gene therapy by using MSCs lentivirally co-transfected with hVEGF165 and hBMP2 for bone regeneration.

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“…Synthetic CT is a therapeutic peptide widely used for treating osteoporosis and CT produces analgesia with a mechanism different from opiate and is effective in patients with refractory pain (Reale et al 2001;Wang et al 2006). However, the major disadvantages of CT therapy are its high cost and low yield.…”

Section: Discussionmentioning

confidence: 99%

“…Two termination codons at the end of the 5hsCT gene were enough to end the translation of the 5hsCT gene (Wang 2005). The Ura3 gene used as an auxotrophic selection marker was equipped with its own promoter and was independently located in front of the expression cassette.…”

Section: Construction Of a Homology-driven Integration Vector With Comentioning

confidence: 99%

“…It encodes 32 amino acids, including 16 amino acids of hCT and 16 amino acids of sCT. The bioactivity of the chimeric calcitonin was higher than hCT (Wang 2005). Next, five copies of hsCT were joined using a segment of DNA encoding a flexible linker, and this construct was incorporated into the expression plasmid pLEX in Escherichia coli GI724.…”

Section: Introductionmentioning

confidence: 99%

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Expression of a chimeric human/salmon calcitonin gene integrated into the Saccharomyces cerevisiae genome using rDNA sequences as recombination sites

Sun

Zang

Liu

et al. 2015

Appl Microbiol Biotechnol

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Calcitonin participates in controlling homeostasis of calcium and phosphorus and plays an important role in bone metabolism. The aim of this study was to endow an industrial strain of Saccharomyces cerevisiae with the ability to express chimeric human/salmon calcitonin (hsCT) without the use of antibiotics. To do so, a homologous recombination plasmid pUC18-rDNA2-ura3-P pgk -5hsCT-rDNA1 was constructed, which contains two segments of ribosomal DNA of 1.1 kb (rDNA1) and 1.4 kb (rDNA2), to integrate the heterologous gene into host rDNA. A DNA fragment containing five copies of a chimeric human/salmon calcitonin gene (5hsCT) under the control of the promoter for phosphoglycerate kinase (P pgk ) was constructed to express 5hsCT in S. cerevisiae using ura3 as a selectable auxotrophic marker gene. After digestion by restriction endonuclease HpaI, a linear fragment, rDNA2-ura3-P pgk -5hsCT-rDNA1, was obtained and transformed into the △ura3 mutant of S. cerevisiae by the lithium acetate method. The ura3-P pgk -5hsCT sequence was introduced into the genome at rDNA sites by homologous recombination, and the recombinant strain YS-5hsCT was obtained. Southern blot analysis revealed that the 5hsCT had been integrated successfully into the genome of S. cerevisiae. The results of Western blot and ELISA confirmed that the 5hsCT protein had been expressed in the recombinant strain YS-5hsCT. The expression level reached 2.04 % of total proteins. S. cerevisiae YS-5hsCT decreased serum calcium in mice by oral administration and even 0.01 g lyophilized S. cerevisiae YS-5hsCT/kg decreased serum calcium by 0.498 mM. This work has produced a commercial yeast strain potentially useful for the treatment of osteoporosis.

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Expression of Human Calcitonin by Microencapsulated Recombinant Myoblasts

Cited by 8 publications

References 13 publications

Clinical significance of metallothioneins in cell therapy and nanomedicine

Clinical significance of metallothioneins in cell therapy and nanomedicine

Osteogenic differentiation effects on rat bone marrow-derived mesenchymal stromal cells by lentivirus-mediated co-transfection of human BMP2 gene and VEGF165 gene

Expression of a chimeric human/salmon calcitonin gene integrated into the Saccharomyces cerevisiae genome using rDNA sequences as recombination sites

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