Abstract 1273: Glioblastoma brain tumor-initiating cells are protected from hypoxia when co-cultured with normal human astrocytes revealing a potential role for mitochondrial transfer via tunneling nanotubes
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Abstract 1273: Glioblastoma brain tumor-initiating cells are protected from hypoxia when co-cultured with normal human astrocytes revealing a potential role for mitochondrial transfer via tunneling nanotubes
Background: Glioblastoma (GBM) has a median survival of Methods: We used a Cytation5 Cell Imager to quantify MU by BTICs from NHAs in direct contact under normal (20%) or hypoxic oxygen tensions (5%). We obtained RT-sensitive (JX14P) patient-derived xenograft BTICs and generated a paired acquired-resistant (JX14P-RT) line. This was achieved by implanting primary tumors into flanks of athymic nude mice and serially treating with 6 fractions of 2Gy over 14 days for multiple passages until the median doubling time was halved. Cells were plated at a 1:1 ratio on Geltrex for 18h and exposed to 5% or 20% oxygen in serum-free media. NHAs were pre-labeled with a GFP-mitochondria tracker and BTICs were infected with a mCherry lentivirus. BTIC-MU was determined by quantifying double-positive cells in whole-well images. Viability was determined using CellTiterGlo, n = 4. Results: Time-lapse imaging revealed GFP-mitochondria transfer from NHAs to BTIC cells via TNTs stimulated by hypoxic conditions. We measured overall MU and cell viability in both BTIC lines. JX14P co-cultured with NHAs trended toward an increased MU (cell fraction) in hypoxia (Hyp) compared to Normoxia (Norm) (Norm = 32.61 ± 13, Hyp = 44.83 ± 5, P>0.167). JX14P exhibit higher cell viability (RLU) in hypoxia when mono- or co-cultured with NHAs (Mono: Norm = 10275 ± 901, Hyp = 12599 ± 579, P0.167). Compared to monoculture, JX14PRT exhibits higher cell viability in co-culture with NHAs under hypoxia (Mono: Norm = 9721 ± 255, Hyp = 10011 ± 1462, P>0.998, Co: Norm = 4928 ± 664, Hyp = 7805 ± 944, P Conclusions: RT-sensitive or -resistant BTICs cocultured with NHAs exhibit increased cell viability under acute hypoxia compared to Normoxia with a trend toward increased MU. Results indicate a potential protective effect following direct interaction with NHAs under hypoxia. We are further exploring metabolic changes in each cell type following mitochondrial exchange. Citation Format: Lauren C. Nassour-Caswell, Nicholas J. Eustace, Christian T. Stackhouse, Hasan Alrefai, Patricia H. Hicks, Taylor L. Schanel, Joshua C. Anderson, Andee M. Beierle, Christopher D. Willey. Glioblastoma brain tumor-initiating cells are protected from hypoxia when co-cultured with normal human astrocytes revealing a potential role for mitochondrial transfer via tunneling nanotubes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1273.
