Dynamic imaging of pancreatic nuclear factor κB (NF-κB) activation in live mice using adeno-associated virus (AAV) infusion and bioluminescence.

Orabi, Abrahim I.; Sah, Swati; Javed, Tanveer A.; Lemon, Kathryn; Good, Misty; Guo, Ping; Xiao, Xianmin; Prasadan, Krishna; Gittes, George K.; Jin, Shunqian; Husain, Sohail Z.

doi:10.1074/jbc.a115.647933

Cited by 8 publications

(9 citation statements)

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“…Luciferase Activity Assay-The interaction between GAS5 or the p27 3Ј-UTR with miR-222 was analyzed as described previously (26). Approximately 400-bp fragments including the putative target region were obtained by PCR.…”

Section: Methodsmentioning

confidence: 99%

Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA

Zheng²,

Mao

et al. 2015

Journal of Biological Chemistry

134

104

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Section: Methodsmentioning

confidence: 99%

Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA

Zheng²,

Mao

et al. 2015

Journal of Biological Chemistry

134

104

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“…Tumor cell growth and invasion are regulated by a network of factors, among which nuclear factor κB (NF-κB) is a critical one that regulates the expression of various genes involved in immunity, stress responses, inflammation and the inhibition of apoptosis, thus providing appropriate conditions for tumor cell progression [22][23][24]. NF-κB is constitutively activated in some tumor cells, including HCC cells, and contributes to the maintenance of the highly proliferative malignant phenotype as well as cellular invasion [22][23][24].…”

Section: Introductionmentioning

confidence: 99%

“…NF-κB is constitutively activated in some tumor cells, including HCC cells, and contributes to the maintenance of the highly proliferative malignant phenotype as well as cellular invasion [22][23][24]. NF-κB regulates several metastasis-related matrix metalloproteinases (MMPs), such as MMP1, MMP3, and MMP9 [25][26][27].…”

Section: Introductionmentioning

confidence: 99%

Isocitrate Dehydrogenase 2 Suppresses the Invasion of Hepatocellular Carcinoma Cells via Matrix Metalloproteinase 9

Tian

Zang

Wang

et al. 2015

Cell Physiol Biochem

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Background/Aims: Isocitrate dehydrogenase 2 (IDH2) is a mitochondrial NADP-dependent isocitrate dehydrogenase, and has been found to be a tumor suppressor in several types of tumors. However, the roles of IDH2 in hepatocellular carcinoma (HCC) as well as underlying mechanisms remain unknown. Methods: The IDH2 and matrix metalloproteinase 9 (MMP9) levels in the specimens from 24 HCC patients were investigated by Western blot and ELISA, respectively. Their relationship was examined by correlation analyses. Patient survival with high IDH2 levels and low IDH2 levels was compared. IDH2 levels and MMP9 levels were modified in a human HCC cell line. The effects of IDH2 or MMP9 modulation on the expression of the other were analyzed. The effects of IDH2 on cell invasion were analyzed in a transwell cell invasion assay. The dependence of nuclear factor κB (NF-κB) signaling was examined using a specific inhibitor. Results: The IDH2 levels significantly decreased in HCC, and were lower in HCC with metastases, compared to those without metastases. IDH2 levels inversely correlated with MMP9 levels in HCC. HCC patients with Low IDH2 had lower 5-year survival. MMP9 levels did not regulate IDH2 levels, while IDH2 inhibited MMP9 levels in HCC cells, in a NF-κB signaling dependent manner, possibly through iκB, to suppress HCC cell invasion. Conclusions: Down regulation of IDH2 may promote HCC cell invasion via NF-κB-dependent increases in MMP9 activity. IDH2 may be a potential therapeutic target for HCC.

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“…The bio-distribution of luciferase expression differed between the constitutive AAV8 SFFV biosensor and the NFκB biosensor. A previous study demonstrated NFκB biosensor activity following administration of AAV by pancreatic infusion in adult mice 21 . Baseline expression rose rapidly in those mice but fell to modest levels by 1 month.…”

Section: Discussionmentioning

confidence: 98%

Generation of light-producing somatic-transgenic mice using adeno-associated virus vectors

Karda

Rahim

Wong

et al. 2020

Sci Rep

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We have previously designed a library of lentiviral vectors to generate somatic-transgenic rodents to monitor signalling pathways in diseased organs using whole-body bioluminescence imaging, in conscious, freely moving rodents. We have now expanded this technology to adeno-associated viral vectors. We first explored bio-distribution by assessing GFP expression after neonatal intravenous delivery of AAV8. We observed widespread gene expression in, central and peripheral nervous system, liver, kidney and skeletal muscle. next, we selected a constitutive SffV promoter and nfκB binding sequence for bioluminescence and biosensor evaluation. An intravenous injection of AAV8 containing firefly luciferase and eGFP under transcriptional control of either element resulted in strong and persistent widespread luciferase expression. A single dose of LPS-induced a 10-fold increase in luciferase expression in AAV8-NFκB mice and immunohistochemistry revealed Gfp expression in cells of astrocytic and neuronal morphology. Importantly, whole-body bioluminescence persisted up to 240 days. We have validated a novel biosensor technology in an AAV system by using an nfκB response element and revealed its potential to monitor signalling pathway in a non-invasive manner in a model of LPS-induced inflammation. This technology complements existing germline-transgenic models and may be applicable to other rodent disease models.Germline light-producing transgenic mice where luciferase expression is controlled by an endogenous promoter, a surrogate promoter or by a minimal promoter downstream of tandem, synthetic, transcription factor binding elements, are used to provide an in vivo readout of physiological and pathological processes 1,2 . One of the advantages of this technology is that every cell will contain a copy of the luciferase transgene and therefore provide a whole-body transgene expression profile under the control of a specific promoter of choice. However, producing germline transgenics requires frequent backcrossing and therefore becomes a time-consuming and costly process, using many rodents.We have previously developed a novel technology which allows the generation of light-producing somatic transgenic rodents, using lentiviral vectors as a proof-of-concept system 3 and have validated this technology both in vitro 4,5 and in vivo 1,6 . We have also demonstrated that signalling pathways in diseased organs can be monitored specifically, continually and in a non-invasive manner 1,2 . Exploiting the immune tolerance of neonatal mice

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Dynamic imaging of pancreatic nuclear factor κB (NF-κB) activation in live mice using adeno-associated virus (AAV) infusion and bioluminescence.

Abstract: Authors are urged to introduce these corrections into any reprints they distribute. Secondary (abstract) services are urged to carry notice of these corrections as prominently as they carried the original abstracts.

Cited by 8 publications

References 0 publications

Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA

Long Non-coding RNA Growth Arrest-specific Transcript 5 (GAS5) Inhibits Liver Fibrogenesis through a Mechanism of Competing Endogenous RNA

Isocitrate Dehydrogenase 2 Suppresses the Invasion of Hepatocellular Carcinoma Cells via Matrix Metalloproteinase 9

Generation of light-producing somatic-transgenic mice using adeno-associated virus vectors

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