TY - JOUR
T1 - Analysis of neuroendocrine clones in NSCLCs using an immuno-guided laser-capture microdissection-based approach
AU - Baldelli, Elisa
AU - Mandarano, Martina
AU - Bellezza, Guido
AU - Petricoin, Emanuel F.
AU - Pierobon, Mariaelena
N1 - Funding Information:
We would like to thank Vienna Ludovini and Jacopo Vannucci for their support with the sample collection and database management and Elisabetta Loreti for her invaluable technical support in preparing the histological sections for the study. This work is dedicated to the patients that participated in the study and to all of those affected by NE transdifferentiated tumors. Conceptualization, E.B. and M.P.; methodology, E.B. M.M. and G.B.; formal analysis, E.B. M.M. and G.B.; resources, E.F.P.; data curation, E.B. M.M. G.B. E.F.P. and M.P.: writing – original draft preparation: E.B. and M.P.; writing – review & editing: M.M. G.B. and E.F.P.; supervision, M.P. All authors have read and agreed to the published version of the manuscript. M.P. is consultant of TheraLink Technologies, Inc. E.F.P. is shareholder and consultant of TheraLink Technologies, Inc. E.F.P. is a shareholder and consultant of Perthera, Inc. None of these relationships played a role in the design, experiments, or data analysis of this study.
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/8/22
Y1 - 2022/8/22
N2 - Clonal evolution and lineage plasticity are key contributors to tumor heterogeneity and response to treatment in cancer. However, capturing signal transduction events in coexisting clones remains challenging from a technical perspective. In this study, we developed and tested a signal-transduction-based workflow to isolate and profile coexisting clones within a complex cellular system like non-small cell lung cancers (NSCLCs). Cooccurring clones were isolated under immunohistochemical guidance using laser-capture microdissection, and cell signaling activation portraits were measured using the reverse-phase protein microarray. To increase the translational potential of this work and capture druggable vulnerabilities within different clones, we measured expression/activation of a panel of key drug targets and downstream substrates of FDA-approved or investigational agents. We isolated intermixed clones, including poorly represented ones (<5% of cells), within the tumor microecology and identified molecular characteristics uniquely attributable to cancer cells that undergo lineage plasticity and neuroendocrine transdifferentiation in NSCLCs.
AB - Clonal evolution and lineage plasticity are key contributors to tumor heterogeneity and response to treatment in cancer. However, capturing signal transduction events in coexisting clones remains challenging from a technical perspective. In this study, we developed and tested a signal-transduction-based workflow to isolate and profile coexisting clones within a complex cellular system like non-small cell lung cancers (NSCLCs). Cooccurring clones were isolated under immunohistochemical guidance using laser-capture microdissection, and cell signaling activation portraits were measured using the reverse-phase protein microarray. To increase the translational potential of this work and capture druggable vulnerabilities within different clones, we measured expression/activation of a panel of key drug targets and downstream substrates of FDA-approved or investigational agents. We isolated intermixed clones, including poorly represented ones (<5% of cells), within the tumor microecology and identified molecular characteristics uniquely attributable to cancer cells that undergo lineage plasticity and neuroendocrine transdifferentiation in NSCLCs.
KW - immuno-guided laser capture microdissection
KW - neuroendocrine transdifferentiation
KW - non-small cell lung cancer
KW - phosphoproteome
KW - reverse-phase protein microarray
KW - signal transduction
UR - http://www.scopus.com/inward/record.url?scp=85136037983&partnerID=8YFLogxK
U2 - 10.1016/j.crmeth.2022.100271
DO - 10.1016/j.crmeth.2022.100271
M3 - Article
C2 - 36046628
AN - SCOPUS:85136037983
SN - 2667-2375
VL - 2
JO - Cell Reports Methods
JF - Cell Reports Methods
IS - 8
M1 - 100271
ER -