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Pam3CSK4, a TLR2 ligand, induces differentiation of glioblastoma stem cells and confers susceptibility to temozolomide

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Summary

Glioblastoma multiforme (GBM) is the most aggressive human brain tumor, and GBM stem cells (GSC) may be responsible for its recurrence and therapeutic resistance. Toll-like receptors (TLRs), which recognize multiple ligands (endogenous and pathogen-associated) and trigger the immune response of mature immune cells, are also expressed by hematopoietic stem and progenitor cells, where their activation results in the differentiation of these cells into myeloid cells. Since TLR expression has been recently described in neural cells, including neural stem cells, we studied TLR expression by GSCs and the effect of stimulation by TLR ligands on promoting GSC differentiation into mature GBM cells. First, our results showed heterogeneous TLR expression by GBM cells from human tumors and, for the first time, by human GSCs defined by their CD133+ and CD44+ phenotypes. Next, the effect of TLR ligands was studied in in vitro cell cultures of neurospheres and CD44+ cells obtained from two GBM cell lines (U-87 and U-118). The expression of GSC markers diminished in the presence of Pam3CSK4 or LPS (TLR2 and TLR4 ligands, respectively), thus indicating TLR-dependent differentiation. Interestingly, simultaneous treatment with Pam3CSK4 plus temozolomide (TMZ), the reference drug in GBM treatment, significantly increased cell death compared to the effect of the ligand alone, which showed no toxicity, or TMZ alone. These results suggest a synergistic effect between Pam3CSK4 and TMZ based on the induction of TLR-dependent GSC differentiation towards mature GBM cells, which exhibited increased sensitivity to chemotherapy, and provide new perspectives in GBM therapy.

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Acknowledgments

This work was supported by the following grants: VLC-Bioclínic-TLR-GBMM-GIL-CERDÁ-2015 from INCLIVA and the University of Valencia, FIS-P114/01669 from the Ministerio de Economía y Competitividad (Instituto de Salud Carlos III) and PROMETEOII/2015/007 from the Generalitat Valenciana. We thank Ms. Ana Clari and Guadalupe Herrera for their technical assistance. The funding sources were not involved in the research and/or preparation of the article.

Funding

The present work was supported by the following grants: VLC-Bioclínic TLR-GBM-GIL-CERDÁ-2015 from INCLIVA and the University of Valencia, FIS-P114/01669 from the Ministerio de Economía y Competitividad (Instituto de Salud Carlos III) and PROMETEOII/2015/007 from the Generalitat Valenciana. The funding sources were not involved in the research and/or preparation of the article.

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Authors and Affiliations

  1. Faculty of Medicine and Odontology, Department of Pathology, University of Valencia, Avenida de Blasco Ibáñez 15, 46010, Valencia, Spain

    Javier Megías, Teresa San-Miguel, Rosario Gil-Benso, Lisandra Muñoz-Hidalgo, David Albert-Bellver, Amara Carratalá, Concha López-Ginés & Miguel Cerdá-Nicolás

  2. Department of Microbiology and Ecology, University of Valencia, Valencia, Spain

    Alba Martínez, Daniel Gozalbo & María Luisa Gil

  3. Estructura de Recerca Interdisciplinar en Biotecnologia i Biomedicina, Universitat de València, Burjasot, Spain

    Alba Martínez, Daniel Gozalbo & María Luisa Gil

  4. Service of Pathology, Clinic Hospital, Valencia, Spain

    Miguel Cerdá-Nicolás

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  1. Javier Megías
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  2. Alba Martínez
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Correspondence to Javier Megías.

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Javier Megías declares that he has no conflicts of interest. Alba Martínez declares that she has no conflicts of interest. Teresa San-Miguel declares that she has no conflicts of interest. Rosario Gil-Benso declares that she has no conflicts of interest. Lisandra Muñoz-Hidalgo declares that she has no conflicts of interest. David Albert-Bellver declares that he has no conflicts of interest. Amara Carratalá declares that she has no conflicts of interest. Daniel Gozalbo declares that he has no conflicts of interest. Concha López-Ginés declares that she has no conflicts of interest. María Luisa Gil declares that she has no conflicts of interest. Miguel Cerdá-Nicolás declares that he has no conflicts of interest.

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Megías, J., Martínez, A., San-Miguel, T. et al. Pam3CSK4, a TLR2 ligand, induces differentiation of glioblastoma stem cells and confers susceptibility to temozolomide. Invest New Drugs 38, 299–310 (2020). https://doi.org/10.1007/s10637-019-00788-2

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