Krüppel-like factor 5 is a crucial mediator of intestinal tumorigenesis in mice harboring combined ApcMin and KRASV12 mutations

<p>Abstract</p> <p>Background</p> <p>Both mutational inactivation of the adenomatous polyposis coli (<it>APC</it>) tumor suppressor gene and activation of the <it>KRAS </it>oncogene are implicated in the pathogenesis of colorectal cancer. Mice harboring a germline <it>Apc</it>^{<it>Min </it>}mutation or intestine-specific expression of the <it>KRAS</it>^<it>V</it>12gene have been developed. Both mouse strains develop spontaneous intestinal tumors, including adenoma and carcinoma, though at a different age. The zinc finger transcription factor Krüppel-like factor 5 (KLF5) has previously been shown to promote proliferation of intestinal epithelial cells and modulate intestinal tumorigenesis. Here we investigated the <it>in vivo </it>effect of <it>Klf5 </it>heterozygosity on the propensity of <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12double transgenic mice to develop intestinal tumors.</p> <p>Results</p> <p>At 12 weeks of age, <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12mice had three times as many intestinal tumors as <it>Apc</it>^{<it>Min </it>}mice. This increase in tumor number was reduced by 92% in triple transgenic <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-mice. The reduction in tumor number in <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-mice was also statistically significant compared to <it>Apc</it>^{<it>Min </it>}mice alone, with a 75% decrease. Compared with <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12, tumors from both <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-and <it>Apc</it>^{<it>Min </it>}mice were smaller. In addition, tumors from <it>Apc</it>^{<it>Min </it>}mice were more distally distributed in the intestine as contrasted by the more proximal distribution in <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12and <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-mice. Klf5 levels in the normal-appearing intestinal mucosa were higher in both <it>Apc</it>^{<it>Min </it>}and <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12mice but were attenuated in <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-mice. The levels of β-catenin, cyclin D1 and Ki-67 were also reduced in the normal-appearing intestinal mucosa of <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-mice when compared to <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12mice. Levels of pMek and pErk1/2 were elevated in the normal-appearing mucosa of <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12mice and modestly reduced in Apc^Min/<it>KRAS</it>^<it>V</it>12/<it>Klf5</it>^+/-mice. Tumor tissues displayed higher levels of both Klf5 and β-catenin, irrespective of the mouse genotype from which tumors were derived.</p> <p>Conclusions</p> <p>Results of the current study confirm the cumulative effect of <it>Apc </it>loss and oncogenic <it>KRAS </it>activation on intestinal tumorigenesis. The drastic reduction in tumor number and size due to <it>Klf5 </it>heterozygosity in <it>Apc</it>^<it>Min</it>/<it>KRAS</it>^<it>V</it>12mice indicate a critical function of KLF5 in modulating intestinal tumor initiation and progression.</p

Krüppel-like factor 5 is essential for proliferation and survival of mouse intestinal epithelial stem cells

Krüppel-like factor 5 (KLF5) is a pro-proliferative transcription factor that is expressed in dividing epithelial cells of the intestinal crypt. Leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5) has been identified as a stem cell marker in both small intestinal and colonic epithelial cells. To determine whether KLF5 regulates proliferation of intestinal stem cells, we investigated the effects of Klf5 deletion specifically from the intestinal stem cells in adult mice. Mice with inducible intestinal stem cell-specific deletion of Klf5 (Lgr5-Klf5fl/fl) were injected with tamoxifen for 5 consecutive days to induce Lgr5-driven Cre expression. Intestinal and colonic tissues were examined by immunohistochemistry at various time points up to 112 days following start of tamoxifen treatment. Klf5 is co-localized in the crypt-based columnar (CBC) cells that express Lgr5. By 11 days following the start of tamoxifen treatment, Lgr5-positive crypts from which Klf5 was deleted exhibited a loss of proliferation that was accompanied by an increase in apoptosis. Beginning at 14 days following the start of tamoxifen treatment, both Klf5 expression and proliferation were re-established in the transit-amplifying epithelial cells but not in the Lgr5-positive CBC cells. By 112 days post-treatment, up to 90% of the Lgr5-positive cells from which Klf5 was deleted were lost from the intestinal crypts. These results indicate a critical role for KLF5 in the survival and maintenance of intestinal stem cells

Aortic stiffness index and its association with cardiovascular functions in children before and after transcatheter closure of PDA

Background: Patent ductus arteriosus is generally associated with hyperdynamic status. Given the vascular shunt between the aorta and pulmonary artery, intrinsic aortic changes occur (aortic stiffness). In the present study, we attempted to assess the impact of PDA on aortic stiffness and its connection with cardiovascular function before and after transcatheter closure of PDA. Patient and methods: Our study consisted of 60 children who were preparing for transcatheter closure of PDA and 60 healthy controls. All patients had clinical and echocardiographic proof of hemodynamically significant PDA. Results: Patients with PDA exhibited significantly higher ASI than controls before closure (p-value < 0.05). After closure, ASI was significantly reduced (p-value < 0.05), but still higher than that of controls (p-value < 0.05) at the six-month follow-up assessment. Patients with PDA had significantly lower LVEF than controls before closure (p-value < 0.05). After closure, LVEF was significantly enhanced (p-value < 0.05), and no significant difference was noted amongst patients and controls (p-value < 0.05) at the six-month follow-up assessment. Conclusion: Aortic stiffness is significantly increased in patients with PDA regardless of PDA size. Aortic stiffness is related to reduced heart function. ASI may be valuable for observing the course of patients with PDA before and after intervention. Keywords: Aortic stiffness index, PDA closure, Left ventricular ejection fractio