Cell Biology

Introduction

Our research group specializes in working with a wide range of cancer cells. In addition to established cell lines such as HeLa, we have successfully cultured numerous primary tumor samples obtained through collaborations with hospitals.

Using a specialized isolation technique, we can extract specific cells from tumor tissues - the so called cancer stem cells (CSCs).

CSCs are a subpopulation of cancer cells with distinct properties that contribute to tumor recurrence, therapy resistance, and metastasis (Fig. 1) [1]. In our research, we have characterized CSCs from various tumor types, including endometrial cancer, glioblastoma, prostate cancer and chondrosarcoma. We analyze their surface marker expression, gene expression profiles, and key signaling pathways to better understand their biology, with a special focus on the role of transcription factor NF-κB (Fig. 2). By identifying cancer-specific characteristics, we can differentiate CSCs from healthy cells, paving the way for safer and more targeted cancer therapies.

Furthermore, we assess CSC sensitivity to various chemotherapeutic agents and NF-κB inhibitors, and also explore their interaction with natural killer (NK) cells in the context of CAR-NK cell therapy. These studies provide valuable insights into the efficacy of specific chemotherapy treatments for different tissue origins and highlight potential avenues for advancing immunotherapy [2,3].

Our cultured CSCs serve as a crucial platform for fundamental medical research. Their human-derived origin makes them a valuable alternative to animal models, helping to refine preclinical testing and improve translational cancer therapies.

Cancer Stem Cells of Lung Origin

Our current research project focuses on the isolation and cultivation of lung cancer stem cells (LCSCs) from squamous cell lung carcinoma. We characterize these LCSCs based on their marker profile using immunocytochemistry (ICC). Additionally, we investigate the activation of NF-κB and its potential interaction with factors in a sex-specific manner and under different conditions of hypoxia and normoxia. A particular focus here is on the transcription factor HIF-1α, which is responsible for the regulation of thousands of genes in the context of the control of oxygen demand and supply to the cell. To further advance our understanding, we employ mass spectrometry-based approaches to identify surface markers of LCSCs. Specific LCSC surface markers are crucial not only for their identification and characterization, but also for their potential as novel targets for immunotherapies against lung cancer.

Cancer Initiation by Gene Mutation

Genetic mutations are frequently linked to cancer initiation, as they can disrupt key regulatory pathways that control cell proliferation and survival. One such critical pathway involves the transcription factor NF-κB, which activity is often found to be altered and highly enriched in various cancer types [4, 5].

In this project, we aim to investigate the role of NF-κB mutation in cancer initiation and progression. In Glioblastoma (GBM), an aggressive brain cancer, NF-κB is increasingly active (Fig. 3) [6]. To further investigate the oncogenic potential of NF-κB mutations, we employ an RNP-based CRISPR-approach to generate targeted gene knockouts in various cell models.

Successfully mutated cells will undergo various procedures and extensive characterization including gene expression profiling, CSC marker analysis, morphological assessments, and survival and behavioral studies in comparison to primary isolated GMB CSCs. These experiments will leverage cutting-edge technologies such as the Mantis liquid dispenser or newest confocal laser scanning microscopy, alongside classical cell culture techniques. By elucidating the impact of NF-κB mutations on tumor initiation, we aim to uncover novel mechanistic insights and identify potential therapeutic targets for more effective cancer treatments.

References

[1] Kaltschmidt et al.: A Role for NF-κB in Organ Specific Cancer and Cancer Stem Cells. Cancers (Basel). 2019.

[2] Helweg et al.: Targeting Key Signaling Pathways in Glioblastoma Stem Cells for the Development of Efficient Chemo- and Immunotherapy. Int J Mol Sci. 2022.

[3] Witte et al.: Nanopore Sequencing Reveals Global Transcriptome Signatures of Mitochondrial and Ribosomal Gene Expressions in Various Human Cancer Stem-like Cell Populations. Cancers (Basel). 2021.

[4] Witte et al.: Nanopore Sequencing Reveals Global Transcriptome Signatures of Mitochondrial and Ribosomal Gene Expressions in Various Human Cancer Stem-like Cell Populations. Cancers. 2021.

[5] Kaltschmidt et al.: Subunit-Specific Role of NF-κB in Cancer. Biomedicines. 2018.

[6] Puliyappadamba et al.: The role of NF-κB in the pathogenesis of glioma. Mol Cell Oncol. 2014.