A Transgenic Model Reveals the Role of Klotho in Pancreatic Cancer Development and Paves the Way for New Klotho-Based Therapy.

Anat Klein-Goldberg,Arkadi Hesin,Miguel Chillón,Hannes Olauson,Assumpció Bosch,Carmela R. Abraham,Ella Zeldich,Kenneth Samuel Hollander,Ido Wolf,Gilad W. Vainer,Tammi Arbel Rubinstein,Tobias E. Larsson,Ayelet Shabtay-Orbach,Tami Rubinek,Inbal Reuveni

doi:10.3390/cancers13246297

Abstract

Simple SummaryWe aimed to study the role of the anti-aging protein klotho and its secreted isoform, sKL, in pancreatic cancer. Three in vivo models, including a novel genetic mouse model and bioinformatics analyses, indicated klotho as a tumor suppressor in pancreatic ductal adenocarcinoma, and unveiled a unique klotho DNA hypermethylation pattern in pancreatic tumors. These results possess significant prognostic value, and further suggest that sKL may serve as a therapeutic agent for pancreatic ductal adenocarcinoma.Klotho is an anti-aging transmembrane protein, which can be shed and can function as a hormone. Accumulating data indicate that klotho is a tumor suppressor in a wide array of malignancies, and designate the subdomain KL1 as the active region of the protein towards this activity. We aimed to study the role of klotho as a tumor suppressor in pancreatic ductal adenocarcinoma (PDAC). Bioinformatics analyses of The Cancer Genome Atlas (TCGA) datasets revealed a correlation between the survival of PDAC patients, levels of klotho expression, and DNA methylation, and demonstrated a unique hypermethylation pattern of klotho in pancreatic tumors. The in vivo effects of klotho and KL1 were examined using three mouse models. Employing a novel genetic model, combining pancreatic klotho knockdown with a mutation in Kras, the lack of klotho contributed to PDAC generation and decreased mousece survival. In a xenograft model, administration of viral particles carrying sKL, a spliced klotho isoform containing the KL1 domain, inhibited pancreatic tumors. Lastly, treatment with soluble sKL prolonged survival of Pdx1-Cre; KrasG12D/+;Trp53R172H/+ (KPC) mice, a model known to recapitulate human PDAC. In conclusion, this study provides evidence that klotho is a tumor suppressor in PDAC. Furthermore, these data suggest that the levels of klotho expression and DNA methylation could have prognostic value in PDAC patients, and that administration of exogenous sKL may serve as a novel therapeutic strategy to treat PDAC.

Highlights

Pancreatic cancer is among the most aggressive cancers, with a 5-year survival rate of9%
Employing a novel genetic model, we showed that pancreatic klotho knockdown contributed to pancreatic ductal adenocarcinoma (PDAC) development, and reduced survival of Kras mutant mice
Examination of methylation data showed a compatible trend of hampered overall survival (OS) and progression-free interval (PFI) in patients who had tumors with high KLOTHO DNA methylation levels, compared to low (n = 45 and n = 46, respectively; Figure 2A,B)

Summary

Introduction

Pancreatic cancer is among the most aggressive cancers, with a 5-year survival rate of9%. Pancreatic cancer is among the most aggressive cancers, with a 5-year survival rate of. Incidence and mortality are rising, with 60,430 new cases and 48,220 deaths expected in 2021 in the US alone [1]. Current models indicate its development from benign pancreatic intraepithelial neoplasia (PanIN) 1–3 to invasive carcinoma, along with acquisition of genetic mutations. Activation of Kras is considered the first step towards malignancy [2], and in mouse models, leads to the development of PanIN by 9 months [3]. Overt PDAC in these mice is rare, in the KPC model, combining Kras mutation with loss of the tumor suppressor p53 [4], PDAC usually develops by 3–6 months of age [5]

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