In vivo adenovirus-mediated gene transduction into mouse endometrial glands: a novel tool to model endometrial cancer in the mouse

Beauparlant, Stephen L.; Read, Peter W.; Cristofano, Antonio Di

doi:10.1016/j.ygyno.2004.06.008

Cited by 15 publications

(12 citation statements)

References 23 publications

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“…Both coxsackie adenovirus receptors (CAR) receptors and integrins are critical for adenoviral infection (Bergelson et al, 1997;Wickham et al, 1993). Thus, our findings are consistent with previous observations of higher expression of CAR as well as αvβ3 and αvβ5 integrins in uterine epithelial cells with only lower levels detected in stromal cells (Aplin et al, 1996;Beauparlant et al, 2004). However, it is also possible that the epithelial basement membrane functions as a barrier in preventing viral particles to travel to the underneath stromal cells.…”

Section: Intraluminal Adenoviral Cre Infusion Induces Conditional Gensupporting

confidence: 92%

Conditional gene recombination by adenovirus-driven Cre in the mouse uterus

Wang

Xie

Zhang

et al. 2006

genesis

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SummaryCre-mediated conditional gene targeting has been shown to be successful in many cell and tissue types. However, gene recombination in the uterus with heterogeneous cell types by Cre activation is not yet well established. Using recombinant adenoviruses expressing a functional Cre (ADVCre) and ROSA26 reporter mice, we show here that ADV-Cre infused intraluminally in a small volume (10 μl) conditionally excises the loxP site, resulting in lacZ expression in uterine luminal epithelial cells without significantly affecting pregnancy. In contrast, a similar intraluminal infusion of ADV-Cre in a larger volume (50 μl) damages the normal architecture and integrity of the luminal epithelium, inducing gene recombination in the underneath stromal cells, with disruption of pregnancy. Further, decidualizing stromal cells at the implantation sites can be targeted by ADV-Cre after intravenous administration on days 5-6. This route of administration also elicits Cre activity in other tissues, including the liver, spleen, ovary, and, more remarkably, in the adrenal cortex. These findings demonstrate the feasibility of achieving conditional expression or deletion of specific genes in uterine cells at desired times and physiological states.

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Section: Intraluminal Adenoviral Cre Infusion Induces Conditional Gensupporting

confidence: 92%

Conditional gene recombination by adenovirus-driven Cre in the mouse uterus

Wang

Xie

Zhang

et al. 2006

genesis

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“…To exclude this possibility and to determine if Lkb1 functions in a cell-autonomous manner, we injected adenovirus expressing the Cre recombinase (Ad-Cre) into the uterine lumen of female mice homozygous for a floxed Lkb1 allele (L). As previously shown, this approach results in efficient and transient transduction of endometrial epithelium but, because of the presence of tight junctions, not of endometrial stromal or other cell types within the uterus (19).…”

Section: Female Lkb1mentioning

confidence: 83%

“…Experimental Lkb1 À/+ animals were generated by outcrossing to FVB mice; Lkb1 L/L mice were propagated as homozygotes. Intrauterine Ad-Cre injections of Lkb1 L/L mice were done as described (19) with modifications: Ad5CMVCre adenovirus (University of Iowa) was stored in 3% sucrose/PBS at 6 Â 10 À10 plaque-forming units (pfu)/AL in 20-AL aliquots, thawed immediately on ice before use, diluted in MEM (Sigma) to a final concentration of 2.4 Â 10 À9 pfu/AL, and incubated at room temperature Â 30 min. Female mice (6-8 weeks old) at diestrus or postestrus were anesthetized, and Genemate 57-mm mulitFlex Tips (ISCBioExpress) connected to 1-mL syringes were used to deliver 50 AL of adenovirus/MEM through the cervical os and into each uterine horn.…”

Section: Methodsmentioning

confidence: 99%

Loss of Lkb1 Provokes Highly Invasive Endometrial Adenocarcinomas

Contreras¹,

Gurumurthy

Haynie³

et al. 2008

Cancer Research

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Mutations in the LKB1 tumor suppressor gene result in the Peutz-Jeghers syndrome, an autosomal dominant condition characterized by hamartomatous polyps of the gastrointestinal tract and a dramatically increased risk of epithelial malignancies at other sites, including the female reproductive tract. Here we show that female mice heterozygous for a null Lkb1 allele spontaneously develop highly invasive endometrial adenocarcinomas. To prove that these lesions were indeed due to Lkb1 inactivation, we introduced an adenoviral Cre vector into the uterine lumen of mice harboring a conditional allele of Lkb1. This endometrial-specific deletion of the Lkb1 gene provoked highly invasive and sometimes metastatic endometrial adenocarcinomas closely resembling those observed in Lkb1 heterozygotes. Tumors were extremely well differentiated and histopathologically distinctive and exhibited alterations in AMP-dependent kinase signaling. Although Lkb1 has been implicated in the establishment of cell polarity, and loss of polarity defines most endometrial cancers, Lkb1-driven endometrial cancers paradoxically exhibit (given their highly invasive phenotype) normal cell polarity and apical differentiation. In human endometrial cancers, Lkb1 expression was inversely correlated with tumor grade and stage, arguing that Lkb1 inactivation or down-regulation also contributes to endometrial cancer progression in women. This study shows that Lkb1 plays an important role in the malignant transformation of endometrium and that Lkb1 loss promotes a highly invasive phenotype. [Cancer Res 2008;68(3):759-66]

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“…All mouse experiments were carried out in accordance with protocols approved by the Institutional Animal Care and Use Committees of Dana-Farber Cancer Institute and Harvard Medical School. Intrauterine adenovirus expression Cre (Ade-Cre) injections were done in female mice at the age of 8-12 weeks as described (23) with the following modifications: 5ul of plaque-forming units (pfu) Ade-Cre (5×10 7 pfu, University of Iowa) with 0.1% Evans blue (Sigma-Aldrich Inc., St. Louis, MO) was injected through the right uterine horn into the lumen while the left uterine horn was uninjected.…”

Section: Methodsmentioning

confidence: 99%

“…However, since PTEN inactivation driven by PR-cre occurs during early embryogenesis (as early as embryonic day 10), and since PR is also expressed in the stroma, this mouse model failed to faithfully mimic sporadic endometrial cancer in humans. In an alternative approach, somatic deletion of individual genes in the endometrium was achieved by delivering adenovirus-expressing Cre into the uterine lumen(23). With use of this method, recent studies in genetic mouse models have shown that somatic deletion of either Pten or Lkb1 specifically in the endometrium induces endometrial tumorigenesis with partial penetrance(20, 24).…”

Section: Introductionmentioning

confidence: 99%

A Genetic Mouse Model of Invasive Endometrial Cancer Driven by Concurrent Loss of Pten and Lkb1 Is Highly Responsive to mTOR Inhibition

et al. 2014

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Signals from the tumor suppressors PTEN and LKB1 converge on mTOR to negatively regulate its function in cancer cells. Notably, both of these suppressors are attenuated in a significant fraction of human endometrial tumors. In this study, we generated a genetic mouse model of endometrial cancer driven by concomitant loss of these suppressors to gain pathophysiological insight into this disease. Dual loss of Pten and Lkb1 in the endometrial epithelium led to rapid development of advanced endometrioid endometrial tumors with 100% penetrance and short host survival. The tumors displayed dysregulated PI3K/Akt and Lkb1/Ampk signaling with hyperactivation of mTOR signaling. Treatment with a dual PI3K/mTOR inhibitor, BEZ235, extended the time before tumor onset and prolonged overall survival. The PI3K inhibitor GDC-0941 used as a single agent reduced the growth rate of primary tumor implants in Pten/Lkb1-deficient mice, and the mTOR inhibitor RAD001 was unexpectedly as effective as BEZ235 in triggering tumor regression. In parallel, we also found that ectopic expression of LKB1 in PTEN/LKB1-deficient human endometrial cancer cells increased their sensitivity to PI3K inhibition. Together, our results demonstrated that Pten/Lkb1-deficient endometrial tumors rely strongly on deregulated mTOR signaling, and they provided evidence that LKB1 status may modulate the response of PTEN-deficient tumors to PI3K or mTOR inhibitors.

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In vivo adenovirus-mediated gene transduction into mouse endometrial glands: a novel tool to model endometrial cancer in the mouse

Cited by 15 publications

References 23 publications

Conditional gene recombination by adenovirus-driven Cre in the mouse uterus

Conditional gene recombination by adenovirus-driven Cre in the mouse uterus

Loss of Lkb1 Provokes Highly Invasive Endometrial Adenocarcinomas

A Genetic Mouse Model of Invasive Endometrial Cancer Driven by Concurrent Loss of Pten and Lkb1 Is Highly Responsive to mTOR Inhibition

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