Tumor necrosis factor receptor 2-signaling in CD133-expressing cells in renal clear cell carcinoma

Abstract

Compared to normal kidney, renal clear cell carcinomas (ccRCC) contain increased numbers of interstitial, non-hematopoietic CD133+cells that express stem cell markers and exhibit low rates of proliferation. These cells fail to form tumors upon transplantation but support tumor formation by differentiated malignant cells. We hypothesized that killing of ccRCC CD133+ (RCCCD133+) cells by cytotoxic agents might be enhanced by inducing them to divide. Since tumor necrosis factor-alpha (TNF), signalling through TNFR2, induces proliferation of malignant renal tubular epithelial cells, we investigated whether TNFR2 might similarly affect RCCCD133+cells. We compared treating organ cultures of ccRCC vs adjacent nontumour kidney (NK) and RCCCD133+ vs NK CD133+ (NKCD133+) cell cultures with wild-type TNF (wtTNF) or TNF muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF). In organ cultures, R2TNF increased expression of TNFR2 and promoted cell cycle entry of both RCCCD133+ and NKCD133+ but effects were greater in RCCCD133+. In contrast, R1TNF increased TNFR1 expression and promoted cell death. Importantly, cyclophosphamide triggered much more cell death in RCCCD133+ and NKCD133+cells pre-treated with R2TNF as compared to untreated controls. We conclude that selective engagement of TNFR2 by TNF can drives RCCCD133+ proliferation and thereby increase sensitivity to cell cycle-dependent cytotoxicity.

Tumor necrosis factor receptor‐2 signaling pathways promote survival of cancer stem‐like CD133 + cells in clear cell renal carcinoma

“…21 Isolated cells were strongly positive for CD133 and exhibited a stem-cell phenotype evidenced by strong immunostaining for SSEA-4 and Nanog ( Figure 1E,F), as previously reported. 21 Isolated cells were strongly positive for CD133 and exhibited a stem-cell phenotype evidenced by strong immunostaining for SSEA-4 and Nanog ( Figure 1E,F), as previously reported.…”

“…21 In brief, ~2 cm 2 tissue pieces were digested into single-cell suspension using Miltenyi Tissue Dissociation Kit on a GentleMACs Disassociator and incubated for 30min (37°C) on a MACsMix rotator, then passed through a 40 μm strainer. 21 In brief, ~2 cm 2 tissue pieces were digested into single-cell suspension using Miltenyi Tissue Dissociation Kit on a GentleMACs Disassociator and incubated for 30min (37°C) on a MACsMix rotator, then passed through a 40 μm strainer.…”

“…15 For STAT3, JAK signaling is typically activated in response to cytokines that bind to receptors containing gp130, inducing the JAK-mediated phosphorylation of tyrosine-705. 21 The current investigation was initiated to understand the mechanism through which TNFR2 acts. Neither TNF receptor has been linked to this response.…”

“…21 For organ cultures, deparaffinized sections were exposed to high-pressure antigen retrieval before incubation with anti-pSTAT3 Ser727 , -CD133 or -TNFR2 (1:100) overnight (4°C). 21 For organ cultures, deparaffinized sections were exposed to high-pressure antigen retrieval before incubation with anti-pSTAT3 Ser727 , -CD133 or -TNFR2 (1:100) overnight (4°C).…”

The features of expression and prognostic significance of the peptide molecules connected with the key properties of renal cell carcinoma

“…[20]. Пред-полагается, что CD133-позитивные клетки в по-чечно-клеточном раке поддерживают клеточную популяцию опухоли и служат источником диффе-ренцированных злокачественных клеток [21].…”