LIF/LIFR oncogenic signaling is a novel therapeutic target in endometrial cancer

Uday P Pratap,Philip T Valente,Sureshkumar Mulampurath Achuthan Pillai ,Kumaraguruparan Ramasamy,Zexuan Liu,Ratna K Vadlamudi,Xiaonan Li,Swapna Konda,Rajeshwar Rao Tekmal ,Bindu Santhamma,Behnam Ebrahimi,Gangadhara R Sareddy,Edward R Kost ,Suryavathi Viswanadhapalli,Logan Blankenship,Prabhakar Pitta Venkata,Kristin A Altwegg,Weiwei Tang,Junhao Liu,Hareesh B Nair

doi:10.1038/s41420-021-00603-z

Abstract

Endometrial cancer (EC) is the fourth most common cancer in women. Advanced-stage EC has limited treatment options with a poor prognosis. There is an unmet need for the identification of actionable drivers for the development of targeted therapies in EC. Leukemia inhibitory factor receptor (LIFR) and its ligand LIF play a major role in cancer progression, metastasis, stemness, and therapy resistance. However, little is known about the functional significance of the LIF/LIFR axis in EC progression. In this study using endometrial tumor tissue arrays, we identified that expression of LIF, LIFR is upregulated in EC. Knockout of LIFR using CRISPR/Cas9 in two different EC cells resulted in a significant reduction of their cell viability and cell survival. In vivo studies demonstrated that LIFR-KO significantly reduced EC xenograft tumor growth. Treatment of established and primary patient-derived EC cells with a novel LIFR inhibitor, EC359 resulted in the reduction of cell viability with an IC50 in the range of 20–100 nM and induction of apoptosis. Further, treatment with EC359 reduced the spheroid formation of EC cancer stem cells and reduced the levels of cancer stem cell markers SOX2, OCT4, NANOG, and Axin2. Mechanistic studies demonstrated that EC359 treatment attenuated the activation of LIF-LIFR driven pathways, including STAT3 and AKT/mTOR signaling in EC cells. Importantly, EC359 treatment resulted in a significant reduction of the growth of EC patient-derived explants ex vivo, EC cell line-derived xenografts, and patient-derived xenografts in vivo. Collectively, our work revealed the oncogenic potential of the LIF/LIFR axis in EC and support the utility of LIFR inhibitor, EC359, as a novel targeted therapy for EC via the inhibition of LIF/LIFR oncogenic signaling.

Highlights

Endometrial cancer (EC) accounts for ~76,000 deaths among women worldwide each year
In this study using endometrial tumor tissue arrays, we identified that expression of Leukemia inhibitory factor (LIF), Leukemia inhibitory factor receptor (LIFR) is upregulated in EC
We have examined the role of LIF/LIFR signaling in 1-A cells were treated with EC359 and the apoptosis was measured

Summary

INTRODUCTION

Endometrial cancer (EC) accounts for ~76,000 deaths among women worldwide each year. It is the sixth leading cause of cancer death among women in the United States [1]. LIF is a commonly upregulated gene in carboplatin- and paclitaxel-resistant cells and its expression correlates with poor outcome in EC patients [23] LIF/LIFR may function as a novel therapeutic target for EC, compromised the activity of EC359 to reduce the viability of both its role and mechanisms in the progression of EC remain elusive. Results showed that EC359 treatment induces apoptosis (Fig. 3F) These findings indicated that EC359 inhibits LIFR and is highly potent in cell lines, we provide genetic evidence that intrinsic LIF/LIFR reducing cell viability, survival, and inducing apoptosis in EC cells. Our work has shown that novel LIFR inhibitor, EC359, reduces the growth of EC cell lines EC359 reduced STAT3 reporter activity and STAT3 target gene with high potency and promotes apoptosis.

RESULTS

DISCUSSION

MATERIALS AND METHODS

Findings

F Control EC359