Pathway‐focused gene expression profiles and immunohistochemistry detection identify contrasting association of caspase 3 (CASP3) expression with prognosis in pediatric classical Hodgkin lymphoma

Abstract

The search for clinically relevant molecular markers in classical Hodgkin lymphoma (cHL) is hampered by the histopathological complexity of the disease, resulting from the admixture of a small number of neoplastic Hodgkin and Reed-Sternberg (H-RS) cells with an abundant and heterogeneous microenvironment. In this study, we evaluated gene expression profiles of 11 selected genes previously proposed as a molecular score for adult cHL, aiming to validate its application in the pediatric setting. Assays were performed by RT-qPCR from formalin-fixed paraffin-embedded (FFPE) lymph nodes in 80 patients with cHL. Selected genes were associated with cell cycle (CENPF, CDK1, CCNA2, CCNE2, and HMMR), apoptosis (BCL2, BCL2L1, and CASP3), and monocytes/macrophages (LYZ and STAT1). Despite using controlled preanalytical and analytical strategies, we were not able to validate the 11-gene score to be applied in pediatric cHL. Principal component analysis (PCA) disclosed 3 components that accounted for 65.7% of the total variability. The second PC included microenvironment and apoptosis genes, from which CASP3 expression was associated with a short time of progression-free survival, which impact was maintained in the unfavorable risk group, Epstein-Barr virus-negative cases, and multivariate analysis (P < .05). Because this is a counterintuitive association, CASP3 active expression was assessed at the protein level in H-RS cells by double immunohistochemistry. In contrast to the association of mRNA levels with a poor therapeutic response, a high number of cleaved CASP3+ cells were associated with longer progression-free survival (P = .03) and overall survival (P = .002). Our results demonstrate the feasibility of using FFPE samples as RNA source for molecular prognostication, but argue against the concept of direct and wide applicability of molecular scores in cHL. We reinforce the potential of CASP3 as an interesting target to be explored in adult and pediatric cHL, and alert for its dual biological role in H-RS cells and tumor microenvironment.

The circuitry of the tumor microenvironment in adult and pediatric Hodgkin lymphoma: cellular composition, cytokine profile, EBV, and exosomes

“… 45 , 46 TGF‐β has been detected predominantly in nodular sclerosis (NS) HL and has potent anti‐inflammatory properties. 35 , 47 Caspase3 (CASP3) is a marker of cell death by apoptosis, and the fact that high levels of CASP3 + H‐RS cells correlate with a better outcome in adult 48 and pediatric 49 , 50 HL underscores its role in apoptosis of H‐RS cells. Thus, CASP3 can serve as a potential biomarker for response prediction of HL.…”

Spatial maps of hepatocellular carcinoma transcriptomes reveal spatial expression patterns in tumor immune microenvironment

“…We examined the spatial expression of marker genes previously reported for common cell types in two samples to assess the sensitivity of the method of detecting the transcripts per spot, and the results confirmed that ALB and CYP2E1 14 were highly expressed in para-carcinoma regions; GPC3 26 and AKR1B10 27 in carcinoma regions; ACTA2 and COL1A1 (markers typically associated with activated fibroblasts or cancer-associated fibroblasts) 28 in the fiber cord and stromal regions, and the above also confirmed the reliability of ST sequencing results. We also detected the spatial distribution of PTPRC (leukocyte marker) 29 , CD2 (T cell and NK cell marker) 30 , and LYZ (myeloid cell marker) 31 in two samples (Figure 1 E); however, no evident spatial characteristics of these markers were found.…”