Surgical resection of glioblastomas induces pleiotrophin-mediated self-renewal of glioblastoma stem cells in recurrent tumors

Arnon Møldrup Knudsen; Bo Halle; Oriane Cédile; Mark Burton; Christina Baun; Helge Thisgaard; Atul Anand; Christopher Hubert; Mads Thomassen; Signe Regner Michaelsen; Birgitte Brinkmann Olsen; Rikke Hedegaard Dahlrot; Rolf Bjerkvig; Justin Durla Lathia; Bjarne Winther Kristensen

doi:10.1093/neuonc/noab302

Surgical resection of glioblastomas induces pleiotrophin-mediated self-renewal of glioblastoma stem cells in recurrent tumors

Neuro Oncol. 2022 Jul 1;24(7):1074-1087. doi: 10.1093/neuonc/noab302.

Authors

Arnon Møldrup Knudsen^{1

2}, Bo Halle^{1

3}, Oriane Cédile⁴, Mark Burton^{5

6}, Christina Baun^{7

8}, Helge Thisgaard^{1

7}, Atul Anand^{1

2}, Christopher Hubert^{9

10

11}, Mads Thomassen^{1

5

6}, Signe Regner Michaelsen¹², Birgitte Brinkmann Olsen^{1

7}, Rikke Hedegaard Dahlrot^{1

13}, Rolf Bjerkvig^{14

15}, Justin Durla Lathia^{10

11

16

17}, Bjarne Winther Kristensen^{1

2

12

18}

Affiliations

¹ Department of Clinical Research, University of Southern Denmark, Odense, Denmark.
² Department of Pathology, Odense University Hospital, Odense, Denmark.
³ Department of Neurosurgery, Odense University Hospital, Odense, Denmark.
⁴ Hematology-Pathology Research Laboratory, Research Unit for Hematology and Research Unit for Pathology, University of Southern Denmark and Odense University Hospital, Odense, Denmark.
⁵ Department of Clinical Genetics, Odense University Hospital, Odense, Denmark.
⁶ Clinical Genome Center, University of Southern Denmark & Region of Southern Denmark, Odense, Denmark.
⁷ Department of Nuclear Medicine, Odense University Hospital, Odense, Denmark.
⁸ Danish Molecular Biomedical Imaging Center (DaMBIC), University of Southern Denmark, Odense, Denmark.
⁹ Department of Biomedical Engineering, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio, USA.
¹⁰ Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, Ohio, USA.
¹¹ Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, USA.
¹² Department of Pathology, Bartholin Institute, Copenhagen University Hospital, Copenhagen, Denmark.
¹³ Department of Oncology, Odense University Hospital, Odense, Denmark.
¹⁴ Department of Biomedicine, University of Bergen, Bergen, Norway.
¹⁵ NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, Strassen, Luxembourg.
¹⁶ Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University, Cleveland, Ohio, USA.
¹⁷ Rose Ella Burkhardt Brain Tumor and Neuro-Oncology Center, Cleveland Clinic, Cleveland, Ohio, USA.
¹⁸ Department of Clinical Medicine and Biotech Research & Innovation Centre (BRIC), University of Copenhagen, Copenhagen, Denmark.

Abstract

Background: Glioblastomas are highly resistant to therapy, and virtually all patients experience tumor recurrence after standard-of-care treatment. Surgical tumor resection is a cornerstone in glioblastoma therapy, but its impact on cellular phenotypes in the local postsurgical microenvironment has yet to be fully elucidated.

Methods: We developed a preclinical orthotopic xenograft tumor resection model in rats with integrated 18F-FET PET/CT imaging. Primary and recurrent tumors were subject to bulk and single-cell RNA sequencing. Differentially expressed genes and pathways were investigated and validated using tissue specimens from the xenograft model, 23 patients with matched primary/recurrent tumors, and a cohort including 190 glioblastoma patients. Functional investigations were performed in vitro with multiple patient-derived cell cultures.

Results: Tumor resection induced microglia/macrophage infiltration, angiogenesis as well as proliferation and upregulation of several stem cell-related genes in recurrent tumor cells. Expression changes of selected genes SOX2, POU3F2, OLIG2, and NOTCH1 were validated at the protein level in xenografts and early recurrent patient tumors. Single-cell transcriptomics revealed the presence of distinct phenotypic cell clusters in recurrent tumors which deviated from clusters found in primary tumors. Recurrent tumors expressed elevated levels of pleiotrophin (PTN), secreted by both tumor cells and tumor-associated microglia/macrophages. Mechanistically, PTN could induce tumor cell proliferation, self-renewal, and the stem cell program. In glioblastoma patients, high PTN expression was associated with poor overall survival and identified as an independent prognostic factor.

Conclusion: Surgical tumor resection is an iatrogenic driver of PTN-mediated self-renewal in glioblastoma tumor cells that promotes therapeutic resistance and tumor recurrence.

Keywords: glioblastoma; pleiotrophin; recurrence; self-renewal; tumor resection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Brain Neoplasms* / drug therapy
Carrier Proteins
Cytokines
Glioblastoma* / genetics
Humans
Neoplasm Recurrence, Local / pathology
Neoplastic Stem Cells / metabolism
Positron Emission Tomography Computed Tomography
Rats
Stem Cells
Tumor Microenvironment

Substances

Carrier Proteins
Cytokines
pleiotrophin