AP09562010 “NF-κB signals activity in myeloid suppressor cells generated from bone marrow cells under physiological and pathological conditions”. Scientific supervisor – Tleulieva R.T., Ph.D.

General concept of the project

Previously thought to be pathologically activated immune regulatory cells contributing to cancer progression, it has been found that myeloid-derived suppressor cells (MDSCs) are a normal component of the physiological inflammatory response, playing a critical role in tissue homeostasis. Aberrant activation of NF-κB signaling has been found in cancer-associated MDSC. However, little is known about the signaling activity of NF-κB in physiologically generated MDSCs. In the proposed project, ex vivo models of MDSC in physiological and pathological environments will be generated to identify possible differences in the activity status of NF-κB family proteins from both canonical and non-canonical signals.

The aim of the project:

To evaluate the signaling activity of NF-κB in MDSCs induced under physiological and pathological conditions, two different biological conditions. Thus, the findings will pave the way for future research on the molecular mechanisms underlying the detrimental role of MDSC in certain diseases.

Project objectives: 

Task 1. Generate MDSC from mouse bone marrow (MBM) cells under two different conditions: physiological and pathological.

Explanation: Ex vivo generation of MDSC in physiological and pathological contexts will help to explore the molecular mechanism that determines the role of MDSC in a given biological context. To create a MDSC in a physiological state: first, a medium conditioned with hepatic stellate cells (HpSC) will be preliminarily prepared by enzymatic dissociation of healthy mouse liver and incubation of individual cells in complete RPMI-based medium. Then, the density gradient portion of the mouse BMC will be seeded into complete RPMI-based media containing GM-CSF and HpSC-conditioned media. To obtain MDSC in a pathological context: BMC will be cultured in a medium supplemented with Ehrlich ascites carcinoma supernatant from an Ehrlich carcinoma mouse. Cell cultures obtained under these conditions will be evaluated phenotypically and functionally using flow cytometry and in vitro suppression assay.

Task 2: To study the signaling activity of NF-κB in MDSCs generated under physiological and pathological conditions.

Explanation: Formation of the p50-50 NF-κB homodimer has been shown to be required for immature myeloid cells to acquire immunosuppressive properties. In addition, activation of other NF-κB signaling subunits in MDSC has been found in cancer. In the proposed project, using western blotting and ELISA-based nuclear translocation analysis, we will study representative homo- and heterodimers of NF-κB family proteins, which belong to both canonical and non-canonical NF-κB signaling, in MDSC cultures. obtained under pathological and physiological conditions.

Results

In this short term project, we have developed both in vitro and in vivo models to generate MDSC. We have successfully developed a model of early liver fibrosis in CD1 mice to create a physiological inflammatory environment in vivo, which in turn is a vital organ for local and systemic induction of MDSC. In parallel, we have developed an in vivo model to generate MDSC in a pathological condition using the well-known Ehrlich subcutaneous carcinoma model.

With regard to the in vitro model, we used the well-known cytokine-induced MDSC model to generate MDSC under physiological conditions. At the same time, we used Ehrlich’s ascites to create MDSC in a pathological condition. In addition, cells derived from in vitro models were phenotypically characterized by flow cytometry analysis.

The members of research group:

Raikhan Tleulieva, Ph.D., Project supervisor. Hirsch index: 5. Total – 47 citations. ORCID: 0000-0001-8897-9986. Scopus ID: 6507202148. WoS ID: N-5634-2017.

Aikyn Kali, MSc, Responsible executive. Hirsch index: 3. Total – 31 citations. ORCID: 0000-0002-5129-1165. Scopus ID: 57195580717. WoS ID: AAF-9781-2020.

Nurshat Abdollah, Ph.D, executor. Hirsch index: 4. Total – 38 citations. ORCID: 0000-0002-4769-7824. Scopus ID: 57194001982. WoS ID: R-2193-2016