Intratumoral Heterogeneity Determines the Expression of mTOR-pathway Proteins in Prostate Cancer
2019-12-11

The aim of this study was to evaluate the expression of mammalian target of rapamycin (mTOR), phosphorylated-mTOR (p-mTOR), and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) in prostate cancer (PCa) in order to assess intratumoral heterogeneity and correlation with clinicopathological parameters. Tissue samples from 115 patients undergoing radical prostatectomy were included in a tissue microarray comprising (A) tissue from the tumor center, (B) malignant border of the tumor, (C) tumor-adjacent benign tissue, and (D) tumor-distant benign prostatic tissue. Immune reactive scores 0-12 were correlated with clinical data in reference to localization. A meta-analysis of studies investigating the association between biochemical recurrence (BCR) and parameters of the mTOR pathway was conducted. Regardless of the location within the tumor, cancer tissue showed higher expression of mTOR, p-mTOR, and 4EB-P1 compared to benign tissue (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M1"><mml:mi>p</mml:mi><mml:mo>&lt;</mml:mo><mml:mn>0.01</mml:mn></mml:math>). Significant differences in expression between tissue samples from groups C and D were observed for mTOR and p-mTOR. When considering expression according to the pathological stage, we observed lower p-mTOR expression in pT3 vs. pT2 (7.9 and 6.3; <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M2"><mml:mi>p</mml:mi><mml:mo>=</mml:mo><mml:mn>0.01</mml:mn></mml:math>). After a median follow-up of 74.5 months (IQR 65.0-84.0), 27 patients (23.47%) developed BCR. Weak staining of mTOR was associated with shorter time to BCR (HR: 2.0; <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M3"><mml:mi>p</mml:mi><mml:mo>=</mml:mo><mml:mn>0.049</mml:mn></mml:math>) after correcting for PSA and T stage. However, a significant association of mTOR expression with BCR was found for specimens from the malignant border of the tumor (B) but not the tumor center (A) (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M4"><mml:mi>p</mml:mi><mml:mo>=</mml:mo><mml:mn>0.0034</mml:mn></mml:math> log rank). In a meta-analysis, we found that the expressions of mTOR (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M5"><mml:mfenced open="(" close=")"><mml:mrow><mml:mtext>RR</mml:mtext></mml:mrow></mml:mfenced><mml:mo>=</mml:mo><mml:mn>0.70</mml:mn></mml:math>; 95% CI 0.43-1.12; <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M6"><mml:mi>p</mml:mi><mml:mo>=</mml:mo><mml:mn>0.13</mml:mn></mml:math>) and 4E-BP1 (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M7"><mml:mfenced open="(" close=")"><mml:mrow><mml:mtext>RR</mml:mtext></mml:mrow></mml:mfenced><mml:mo>=</mml:mo><mml:mn>0.86</mml:mn></mml:math>; <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M8"><mml:mi>p</mml:mi><mml:mo>=</mml:mo><mml:mn>0.53</mml:mn></mml:math>) were not statistically associated with BCR, while strong staining of p-mTOR was associated with a lower risk of BCR (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M9"><mml:mfenced open="(" close=")"><mml:mrow><mml:mtext>RR</mml:mtext></mml:mrow></mml:mfenced><mml:mo>=</mml:mo><mml:mn>0.57</mml:mn></mml:math>; <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M10"><mml:mi>p</mml:mi><mml:mo>=</mml:mo><mml:mn>0.002</mml:mn></mml:math>). All 3 markers showed stronger expression in PCa and exhibited local gradients in relation to the border of tumor and healthy tissue. Our results suggest an important role of intratumor heterogeneity for the use of mTOR parameters as biomarkers in PCa.