Retinoblastoma Is Characterized by a Cold, CD8+ Cell Poor, PD-L1− Microenvironment, Which Turns Into Hot, CD8+ Cell Rich, PD-L1+ After Chemotherapy

Miracco, Clelia; Toti, Paolo; Gelmi, Maria Chiara; Aversa, Sara; Baldino, Gennaro; Galluzzi, Paolo; Francesco, Sonia De; Petrelli, Federica; Sorrentino, Ester; Belmonte, Giuseppe; Galimberti, Daniela; Bracco, Sandra; Hadjistilianou, Theodora

doi:10.1167/iovs.62.2.6

Cited by 9 publications

(12 citation statements)

References 55 publications

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“…We observed that previous treatments did not modify the M2‐like polarized CD163 + environment of retinoblastoma, which is in contrast with a recent study of the retinoblastoma immune environment by Miracco et al [8]. They reported a significantly higher number of CD163 + M2‐like TAMs in previously treated human retinoblastoma eyes, compared to eyes enucleated without previous treatment [8].…”

Section: Discussioncontrasting

confidence: 99%

“…Because IFN‐γ‐secreting cells, and specifically CD8 + lymphocytes, were moderately abundant in a high proportion of the retinoblastoma samples, we propose that tumor microenvironment inhibits such CD8 + cells, leading to a decreased level of IFN‐γ secretion [41]. The work by Miracco et al , in contrast, suggests that chemotherapy treatments induce a proinflammatory environment, leading to higher expression of PD‐L1 [8], but our results and others do not confirm their observation [9]. We suggest that M2‐like TAMs and Tregs suppress the activity of CD4 + and CD8 + TILs in retinoblastomas, because they inhibit the production of several proinflammatory cytokines such as IFN‐γ and IL‐2 [42].…”

Section: Discussionmentioning

confidence: 99%

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Identification of immunosuppressive factors in retinoblastoma cell secretomes and aqueous humor from patients

Cuadrado‐Vilanova

Liu

Paco

et al. 2022

The Journal of Pathology

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The microenvironment of retinoblastoma, the solid malignancy of the developing retina, is immunosuppressive. To study the interactions between tumor‐associated microglia/macrophages (TAMs) and tumor cells in retinoblastomas, we analyzed immunohistochemistry markers in 23 patient samples and characterized 105 secreted cytokines of 11 retinoblastoma cell models in culture. We detected profuse infiltration of CD163+ protumoral M2‐like polarized TAMs in eyes enucleated due to cancer progression. Previous treatment of patients increased the number of TAMs but did not affect M2‐like polarization. M2‐like microglia/macrophages were almost absent in five eyes obtained from children enucleated due to nontumoral causes. CD8+ tumor‐infiltrating lymphocytes (TILs) were moderately abundant in tumor eyes and very scarce in nontumoral ones. The expression of the immune checkpoint molecule PD‐L1 was absent in 95% of the tumor samples, which is concordant with the finding of FOXP3+ Tregs infiltrating tumors. We confirmed the pathology results using single‐cell transcriptome analysis of one tumor. We identified the cytokines extracellular matrix metalloproteinase inducer (EMMPRIN) and macrophage migration inhibitory factor (MIF), both with reported immunosuppressive activity, secreted at high levels in retinoblastoma primary cell cultures. Gene expression analysis of a large retinoblastoma cohort and single‐cell transcriptome analysis confirmed that MIF and EMMPRIN were significantly upregulated in retinoblastomas, which led us to quantify both proteins by immunoassays in liquid biopsies (aqueous humor obtained from more than 20 retinoblastoma patients). We found a significant increase in the concentration of MIF and EMMPRIN in cancer patients, compared to 12 noncancer ones. Finally, we showed that macrophages derived from peripheral blood mononuclear cells increased the expression of markers of M2‐like polarization upon exposure to retinoblastoma‐conditioned medium or recombinant MIF. Overall, our findings suggest that retinoblastoma cell secretions induce the protumoral phenotype of this tumor. Our results might have clinical impact in the fields of biomarkers and treatment. © 2022 The Pathological Society of Great Britain and Ireland.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Identification of immunosuppressive factors in retinoblastoma cell secretomes and aqueous humor from patients

Cuadrado‐Vilanova

Liu

Paco

et al. 2022

The Journal of Pathology

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“…Our study showed a few lymphocytes, particularly T cells, in RB. This was consistent with recent studies also showing the absence of CD8+ cells, including T cells, B cells, in RB [ 55 ]. These results suggested that RB has relatively poor immunogenicity compared with most of tumors, which may contribute to the poor efficacy of immunotherapy in RB.…”

Section: Discussionsupporting

confidence: 94%

Single-cell characterization of malignant phenotypes and microenvironment alteration in retinoblastoma

Yang

Xiao

et al. 2022

Cell Death Dis

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Retinoblastoma (RB) is the most common primary intraocular malignancy of childhood. It is known that the tumor microenvironment (TME) regulates tumorigenesis and metastasis. However, how the malignant progression in RB is determined by the heterogeneity of tumor cells and TME remains uncharacterized. Here, we conducted integrative single-cell transcriptome and whole-exome sequencing analysis of RB patients with detailed pathological and clinical measurements. By single-cell transcriptomic sequencing, we profiled around 70,000 cells from tumor samples of seven RB patients. We identified that the major cell types in RB were cone precursor-like (CP-like) and MKI67+ cone precursor (MKI67+ CP) cells. By integrating copy number variation (CNV) analysis, we found that RB samples had large clonal heterogeneity, where the malignant MKI67+ CP cells had significantly larger copy number changes. Enrichment analysis revealed that the conversion of CP-like to MKI67+ CP resulted in the loss of photoreceptor function and increased cell proliferation ability. The TME in RB was composed of tumor-associated macrophages (TAMs), astrocyte-like, and cancer-associated fibroblasts (CAFs). Particularly, during the invasion process, TAMs created an immunosuppressive environment, in which the proportion of TAMs decreased, M1-type macrophage was lost, and the TAMs-related immune functions were depressed. Finally, we identified that TAMs regulated tumor cells through GRN and MIF signaling pathways, while TAMs self-regulated through inhibition of CCL and GALECTIN signaling pathways during the invasion process. Altogether, our study creates a detailed transcriptomic map of RB with single-cell characterization of malignant phenotypes and provides novel molecular insights into the occurrence and progression of RB.

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“…Targeting cancer stem cells have been recognized as an effective strategy for cancer therapy [2]. Retinoblastoma is the most common eye tumor in children that develops from the immature cells of retina [3]; the incidence of this disease is 1 in 15,000-20,000 live births and the median age at rst diagnosis is two years [4]. So far, non-invasive therapeutic strategies for retinoblastoma are limited; current therapeutic methods, including surgery (enucleation) and radiation, usually results in blindness and inactive treatment causes it life-threatening [5].…”

Section: Introductionmentioning

confidence: 99%

SOX2 maintains the stemness of retinoblastoma stem-like cells through Hippo/YAP signaling pathway

Zhao

Zhou

Lü

et al. 2022

Experimental Eye Research

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Background: Cancer stem cells are responsible for tumor initiation and progression in various types of cancer, while, although the existence of retinoblastoma stem cells had been reported, the mechanisms supporting retinoblastoma stemness are poorly understood. In this study, a modi ed method for isolating retinoblastoma stem-like cells for mechanistic study was rst established and an important mechanism underlying SOX2-drived retinoblastoma stemness was subsequently revealed.Methods: The retinoblastoma stem-like cells were isolated by single cell cloning in combination of examination of sphere-forming capacities. The stemness of isolated retinoblastoma stem-like cells were characterized by sphere-forming capacities and the expression of cancer stem cell markers. The SOX2 gene was overexpressed or knocked down by lentivirus system. The transcriptional regulation was identi ed by qRT-PCR, luciferase reporter, nuclear run-on and DNA pull down assay. Spearman analysis was employed for correlation analysis of genes in tumor tissues of retinoblastoma patients.Results: The isolated retinoblastoma stem-like cells exhibited signi cantly enhanced sphere-forming capacity and constantly higher levels of CD44, ABCG2, SOX2 and PAX6, but not CD133. SOX2 positively regulated the stemness of retinoblastoma stem-like cells as identi ed by gene manipulation technology. SOX2 directly binds to the promoters of WWTR1 and YAP1, transcriptionally activates WWTR1 and YAP1, and thereby activating Hippo/YAP signaling. Knockdown of WWTR1 or YAP1 partially abolished the effect of SOX2 on the stemness of retinoblastoma stem-like cells.Conclusion: An effective method for isolation of retinoblastoma stem-like cells was established. The mechanistic study demonstrated that SOX2, as a key deriver, maintains retinoblastoma stemness by activating Hippo/YAP signaling. Inhibition of Hippo/YAP signaling would be an effective strategy for human retinoblastoma caused by aberrant upregulation of SOX2.

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Retinoblastoma Is Characterized by a Cold, CD8+ Cell Poor, PD-L1− Microenvironment, Which Turns Into Hot, CD8+ Cell Rich, PD-L1+ After Chemotherapy

Cited by 9 publications

References 55 publications

Identification of immunosuppressive factors in retinoblastoma cell secretomes and aqueous humor from patients

Identification of immunosuppressive factors in retinoblastoma cell secretomes and aqueous humor from patients

Single-cell characterization of malignant phenotypes and microenvironment alteration in retinoblastoma

SOX2 maintains the stemness of retinoblastoma stem-like cells through Hippo/YAP signaling pathway

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