image analysis to obtain prognostic information for tumor-infiltrating immune cells in hepatocellular carcinoma (hcc)

Prognostic Significance of Immune Cell Infiltration in Hepatocellular Carcinoma

The spatial organization and abundance of tumor-infiltrating immune cells have been proven to impact survival in different cancers, hepatocellular carcinoma (HCC) included1,2,3,4. The prognostic significance of tumor-infiltrating lymphocytes in cancers was first shown on hematoxylin and eosin (H&#38;E) stained sections5,6. Then, a pioneer study by Galon et al. using immunohistochemistry (IHC) demonstrated associations of the densities of CD3+ and CD8+ T cells in colon cancer tissue with prognosis4.
IHC is a gold standard for visualizing, quantifying, and mapping immune cells within tumor tissue for further association with clinical outcomes7. IHC presents several advantages, such as low cost, widespread availability, and compatibility with formalin-fixed paraffin-embedded (FFPE) tissue8. However, accurate assessment of IHC-stained immune cells is a big challenge. Traditional scoring in selected microscopic fields of view is time-consuming and no longer sufficient to ensure the high-quality, reproducible, objective analysis essential for candidate biomarker selection and reliable clinical correlation9. Whole-slide scans can be assessed as a whole image or after subsampling10.
Computerized quantitative assessment of the abundance of immune cells in tissue samples can ensure practical, accurate, reliable, and clinically relevant data11. QuPath is a free, open-source software that enables the digital image analysis of IHC-stained slides and maximizes the amount of information yielded from individual samples12.
Since immature dendritic cells (iDCs), natural killer (NK) cells, and mast cells are involved in anti-tumor immune responses and were shown to correlate with patients&#39; outcomes13,14,15, we here present a step-by-step protocol for assessing their spatial distribution in FFPE HCC tissue and exploring their prognostic impact. CD1a is expressed mainly on the membrane of immature dendritic cells, the density of which has been associated with clinical outcomes in a variety of human tumors16,17. Mast cells can play a protumor or anti-tumor role within the tumor microenvironment (TME)18. They can support angiogenesis and facilitate metastasis19. Conversely, it has been reported that mast cells can mediate tumor cell apoptosis through the production of IL-420. Anti-CD117 staining is commonly used to visualize and quantify mast cells within the tumor tissue21. NK cells are thought to contribute to the surveillance and control of HCC22 by killing cancer cells23. NKp46 expression by NK is a crucial parameter for their anti-tumor activity24. However, NKp46+ NK cells correlated positively with tumor diameter in HCC patients25. Despite this, little is known about their correlation with patient survival.
We aimed to quantitatively assess the abundance of CD1a+ iDCs, CD117+ mast cells, and NKp46+ NK cells in different regions of HCC and highlight their prognostic significance. A total of 70 consecutive patients with pathologically confirmed stage I-IV HCC, who were eligible for resection according to BCLC guidelines and underwent curative-intent liver resection at Pilsen University Hospital between 1997 and 2019, were included in the current retrospective study. Pathology reports of the patients were reviewed. None of the patients included in this study had distant metastases nor had received neoadjuvant therapy such as chemotherapy or radiotherapy before surgery. A total of 3 patients with poor-quality histological specimens were excluded, and the remaining 67 patients were included in the study (Table 1).

Author Spotlight: Investigating Immune Cell Dynamics in the Tumor Microenvironment &amp;#8212; Challenges and Innovations in Cancer Prognosis

Mast Cells in the Microenvironment of Hepatocellular Carcinoma Confer Favorable Prognosis: A Retrospective Study using QuPath Image Analysis Software

Image Analysis to Obtain Prognostic Information for Tumor-Infiltrating Immune Cells in Hepatocellular Carcinoma (HCC)

image-analysis-to-obtain-prognostic-information-for-tumor

Research

JoVE Journal

Cancer Research

216 Views.  Charles University. Image Analysis to Obtain Prognostic Information for Tumor-Infiltrating Immune Cells in Hepatocellular Carcinoma (HCC)

Video: Image Analysis to Obtain Prognostic Information for Tumor-Infiltrating Immune Cells in Hepatocellular Carcinoma (HCC)

The insights provided by in-situ detection of immune cells within hepatocellular carcinoma (HCC) might present information on patient outcomes. Studies investigating the expression and localization of immune cells within tumor tissues are associated with several challenges, including a lack of precise annotation for tumor regions and random selection of microscopic fields of view. QuPath is an open-source, user-friendly software that could meet the growing need for digital pathology in whole-slide image (WSI) analysis.
The infiltration of HCC and adjacent tissues by CD1a+ immature dendritic cells (iDCs), CD117+ mast cells, and NKp46+ natural killer cells (NKs) cells was assessed immunohistochemically in representative specimens of 67 patients with HCC who underwent curative resection. The area fraction (AF) of positively stained cells was assessed automatically in WSIs using QuPath in the tumor center (TC), inner margin (IM), outer margin (OM), and peritumor (PT) area. The prognostic significance of immune cells was evaluated for time to recurrence (TTR), disease-free survival (DFS), and overall survival (OS).
The AF of mast cells was significantly greater than the AF of NKs, and the AF of iDCs was significantly lower compared to NKs in each region of interest. High AFs of mast cells in the IM and PT areas were associated with longer DFS. In addition, high AF of mast cells in IM was associated with longer OS.
Computer-assisted analysis using this software is a suitable tool for obtaining prognostic information for tumor-infiltrating immune cells (iDCs, mast cells, and NKs) in different regions of HCC after resection. Mast cells displayed the greatest AF in all regions of interest (ROIs). Mast cells in the peritumor region and IM showed a positive prognostic significance.

The presence of mast cells in the inner margin and peritumor areas of hepatocellular carcinoma after resection confers a favorable prognosis. This study endorses QuPath image analysis software as a promising platform that could meet the need for reproducibility, consistency, and accuracy in digital pathology.

The insights provided by in-situ detection of immune cells within hepatocellular carcinoma (HCC) might present information on patient outcomes. Studies ...