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Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants

BACKGROUND: High-risk neuroblastomas (HR-NBs) are rare, aggressive pediatric cancers characterized by resistance to therapy and relapse in more than 30% of cases, despite using an aggressive therapeutic protocol including targeting of GD2. The mechanisms responsible for therapy resistance are unclea...

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Autores principales: Dondero, Alessandra, Morini, Martina, Cangelosi, Davide, Mazzocco, Katia, Serra, Martina, Spaggiari, Grazia Maria, Rotta, Gianluca, Tondo, Annalisa, Locatelli, Franco, Castellano, Aurora, Scuderi, Francesca, Sementa, Angela Rita, Eva, Alessandra, Conte, Massimo, Garaventa, Alberto, Bottino, Cristina, Castriconi, Roberta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8021887/
https://www.ncbi.nlm.nih.gov/pubmed/33795387
http://dx.doi.org/10.1136/jitc-2020-002293
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author Dondero, Alessandra
Morini, Martina
Cangelosi, Davide
Mazzocco, Katia
Serra, Martina
Spaggiari, Grazia Maria
Rotta, Gianluca
Tondo, Annalisa
Locatelli, Franco
Castellano, Aurora
Scuderi, Francesca
Sementa, Angela Rita
Eva, Alessandra
Conte, Massimo
Garaventa, Alberto
Bottino, Cristina
Castriconi, Roberta
author_facet Dondero, Alessandra
Morini, Martina
Cangelosi, Davide
Mazzocco, Katia
Serra, Martina
Spaggiari, Grazia Maria
Rotta, Gianluca
Tondo, Annalisa
Locatelli, Franco
Castellano, Aurora
Scuderi, Francesca
Sementa, Angela Rita
Eva, Alessandra
Conte, Massimo
Garaventa, Alberto
Bottino, Cristina
Castriconi, Roberta
author_sort Dondero, Alessandra
collection PubMed
description BACKGROUND: High-risk neuroblastomas (HR-NBs) are rare, aggressive pediatric cancers characterized by resistance to therapy and relapse in more than 30% of cases, despite using an aggressive therapeutic protocol including targeting of GD2. The mechanisms responsible for therapy resistance are unclear and might include the presence of GD2neg/low NB variants and/or the expression of immune checkpoint ligands such as B7-H3. METHOD: Here, we describe a multiparametric flow cytometry (MFC) combining the acquisition of 10(6) nucleated singlets, Syto16pos CD45neg CD56pos cells, and the analysis of GD2 and B7-H3 surface expression. 41 bone marrow (BM) aspirates from 25 patients with NB, at the onset or relapse, are analyzed, comparing results with cytomorphological analysis (CA) and/or immunohistochemistry (IHC). Spike in experiments assesses the sensitivity of MFC. Kaplan-Meier analysis on 498 primary NBs selects novel prognostic markers possibly integrating the MFC panel. RESULTS: No false positive are detected, and MFC shows high sensitivity (0.0005%). Optimized MFC identifies CD45negCD56pos NB cells in 11 out of 12 (91.6%) of BM indicated as infiltrated by CA, 7 of which coexpress high levels of GD2 and B7-H3. MFC detects CD45negCD56posGD2neg/low NB variants expressing high surface levels of B7-H3 in two patients with HR-NB (stage M) diagnosed at 53 and 139 months of age. One of them has a non-MYCN amplified tumor with unusual THpos PHOX2Bneg phenotype, which relapsed 141 months post-diagnosis with BM infiltration and a humerus lesion. All GD2neg/low NB variants are detected in patients at relapse. Kaplan-Meier analysis highlights an interesting dichotomous prognostic value of MML5, ULBPs, PVR, B7-H6, and CD47, ligands involved in NB recognition by the immune system. CONCLUSIONS: Our study validates a sensitive MFC analysis providing information on GD2 and B7-H3 surface expression and allowing fast, specific and sensitive evaluation of BM tumor burden. With other routinely used diagnostic and prognostic tools, MFC can improve diagnosis, prognosis, orienting novel personalized treatments in patients with GD2low/neg NB, who might benefit from innovative therapies combining B7-H3 targeting.
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spelling pubmed-80218872021-04-21 Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants Dondero, Alessandra Morini, Martina Cangelosi, Davide Mazzocco, Katia Serra, Martina Spaggiari, Grazia Maria Rotta, Gianluca Tondo, Annalisa Locatelli, Franco Castellano, Aurora Scuderi, Francesca Sementa, Angela Rita Eva, Alessandra Conte, Massimo Garaventa, Alberto Bottino, Cristina Castriconi, Roberta J Immunother Cancer Immunotherapy Biomarkers BACKGROUND: High-risk neuroblastomas (HR-NBs) are rare, aggressive pediatric cancers characterized by resistance to therapy and relapse in more than 30% of cases, despite using an aggressive therapeutic protocol including targeting of GD2. The mechanisms responsible for therapy resistance are unclear and might include the presence of GD2neg/low NB variants and/or the expression of immune checkpoint ligands such as B7-H3. METHOD: Here, we describe a multiparametric flow cytometry (MFC) combining the acquisition of 10(6) nucleated singlets, Syto16pos CD45neg CD56pos cells, and the analysis of GD2 and B7-H3 surface expression. 41 bone marrow (BM) aspirates from 25 patients with NB, at the onset or relapse, are analyzed, comparing results with cytomorphological analysis (CA) and/or immunohistochemistry (IHC). Spike in experiments assesses the sensitivity of MFC. Kaplan-Meier analysis on 498 primary NBs selects novel prognostic markers possibly integrating the MFC panel. RESULTS: No false positive are detected, and MFC shows high sensitivity (0.0005%). Optimized MFC identifies CD45negCD56pos NB cells in 11 out of 12 (91.6%) of BM indicated as infiltrated by CA, 7 of which coexpress high levels of GD2 and B7-H3. MFC detects CD45negCD56posGD2neg/low NB variants expressing high surface levels of B7-H3 in two patients with HR-NB (stage M) diagnosed at 53 and 139 months of age. One of them has a non-MYCN amplified tumor with unusual THpos PHOX2Bneg phenotype, which relapsed 141 months post-diagnosis with BM infiltration and a humerus lesion. All GD2neg/low NB variants are detected in patients at relapse. Kaplan-Meier analysis highlights an interesting dichotomous prognostic value of MML5, ULBPs, PVR, B7-H6, and CD47, ligands involved in NB recognition by the immune system. CONCLUSIONS: Our study validates a sensitive MFC analysis providing information on GD2 and B7-H3 surface expression and allowing fast, specific and sensitive evaluation of BM tumor burden. With other routinely used diagnostic and prognostic tools, MFC can improve diagnosis, prognosis, orienting novel personalized treatments in patients with GD2low/neg NB, who might benefit from innovative therapies combining B7-H3 targeting. BMJ Publishing Group 2021-04-01 /pmc/articles/PMC8021887/ /pubmed/33795387 http://dx.doi.org/10.1136/jitc-2020-002293 Text en © Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. http://creativecommons.org/licenses/by-nc/4.0/ http://creativecommons.org/licenses/by-nc/4.0/This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Immunotherapy Biomarkers
Dondero, Alessandra
Morini, Martina
Cangelosi, Davide
Mazzocco, Katia
Serra, Martina
Spaggiari, Grazia Maria
Rotta, Gianluca
Tondo, Annalisa
Locatelli, Franco
Castellano, Aurora
Scuderi, Francesca
Sementa, Angela Rita
Eva, Alessandra
Conte, Massimo
Garaventa, Alberto
Bottino, Cristina
Castriconi, Roberta
Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants
title Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants
title_full Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants
title_fullStr Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants
title_full_unstemmed Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants
title_short Multiparametric flow cytometry highlights B7-H3 as a novel diagnostic/therapeutic target in GD2neg/low neuroblastoma variants
title_sort multiparametric flow cytometry highlights b7-h3 as a novel diagnostic/therapeutic target in gd2neg/low neuroblastoma variants
topic Immunotherapy Biomarkers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8021887/
https://www.ncbi.nlm.nih.gov/pubmed/33795387
http://dx.doi.org/10.1136/jitc-2020-002293
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