Long-term adaptation of lymphoma cell lines to hypoxia is mediated by diverse molecular mechanisms that are targetable with specific inhibitors.

in Cell death discovery by Lenka Daumova, Dmitry Manakov, Jiri Petrak, Dana Sovilj, Matej Behounek, Ladislav Andera, Ondrej Vit, Olga Souckova, Ondrej Havranek, Alex Dolnikova, Nicol Renesova, Liliana Tuskova, Lucie Winkowska, Nardjas Bettazova, Kristyna Kupcova, Marie Hubalek Kalbacova, Miriama Sikorova, Marek Trneny, Pavel Klener

TLDR

  • Long-term hypoxia adaptation in NHL cells is associated with decreased proliferation, altered metabolism, and increased dependence on BCL-XL and PI3K-AKT signaling, suggesting potential therapeutic targets.

Abstract

A large body of evidence suggests that hypoxia drives aggressive molecular features of malignant cells irrespective of cancer type. Non-Hodgkin lymphomas (NHL) are the most common hematologic malignancies characterized by frequent involvement of diverse hypoxic microenvironments. We studied the impact of long-term deep hypoxia (1% O2) on the biology of lymphoma cells. Only 2 out of 6 tested cell lines (Ramos, and HBL2) survived ≥ 4 weeks under hypoxia. The hypoxia-adapted (HA)b Ramos and HBL2 cells had a decreased proliferation rate accompanied by significant suppression of both oxidative phosphorylation and glycolytic pathways. Transcriptome and proteome analyses revealed marked downregulation of genes and proteins of the mitochondrial respiration complexes I and IV, and mitochondrial ribosomal proteins. Despite the observed suppression of glycolysis, the proteome analysis of both HA cell lines showed upregulation of several proteins involved in the regulation of glucose utilization including the active catalytic component of prolyl-4-hydroxylase P4HA1, an important druggable oncogene. HA cell lines demonstrated increased transcription of key regulators of auto-/mitophagy, e.g., neuritin, BCL2 interacting protein 3 (BNIP3), BNIP3-like protein, and BNIP3 pseudogene. Adaptation to hypoxia was further associated with deregulation of apoptosis, namely upregulation of BCL2L1/BCL-XL, overexpression of BCL2L11/BIM, increased binding of BIM to BCL-XL, and significantly increased sensitivity of both HA cell lines to A1155463, a BCL-XL inhibitor. Finally, in both HA cell lines AKT kinase was hyperphosphorylated and the cells showed increased sensitivity to copanlisib, a pan-PI3K inhibitor. In conclusion, our data report on several shared mechanisms of lymphoma cell adaptation to long-term hypoxia including: 1. Upregulation of proteins responsible for glucose utilization, 2. Degradation of mitochondrial proteins for potential mitochondrial recycling (by mitophagy), and 3. Increased dependence on BCL-XL and PI3K-AKT signaling for survival. In translation, inhibition of glycolysis, BCL-XL, or PI3K-AKT cascade may result in targeted elimination of HA lymphoma cells.

Overview

  • The study investigates the impact of long-term deep hypoxia (1% O2) on the biology of non-Hodgkin lymphoma (NHL) cells.
  • Researchers exposed 6 NHL cell lines to hypoxia and identified only 2 cell lines (Ramos and HBL2) that survived ≥ 4 weeks.
  • The study aimed to identify common mechanisms by which NHL cells adapt to hypoxia and potentially target these mechanisms for treatment.

Comparative Analysis & Findings

  • The hypoxia-adapted (HA) Ramos and HBL2 cells showed decreased proliferation rates accompanied by suppressed oxidative phosphorylation and glycolytic pathways.
  • Transcriptome and proteome analyses revealed downregulation of genes and proteins involved in mitochondrial respiration complexes I and IV, and mitochondrial ribosomal proteins, as well as upregulation of proteins involved in glucose utilization.
  • The HA cell lines demonstrated increased transcription of key regulators of auto-/mitophagy, and deregulation of apoptosis.

Implications and Future Directions

  • The study suggests that inhibitors of glycolysis, BCL-XL, and PI3K-AKT cascade may be effective treatments for NHL cells adapted to hypoxia.
  • Future studies should investigate the specificity and efficacy of these inhibitors in different NHL subtypes and clinical trials.
  • Understanding the molecular mechanisms of hypoxia adaptation may also lead to the development of more effective hypoxia-targeted therapies.
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