An Experimental and Computational Protocol to Study Cell Proliferation in Human Acute Myeloid Leukemia Xenografts

Thalia Vlachou, Marco S. Nobile, Chiara Ronchini, Daniela Besozzi, Pier Giuseppe Pelicci

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

Acute myeloid leukemia (AML) is a highly frequent hematological malignancy, characterized by clinical and biological diversity, along with high relapse and mortality rates. The inherent functional and genetic intra-tumor heterogeneity in AML is thought to play an important role in disease recurrence and resistance to chemotherapy. Patient-derived xenograft (PDX) models preserve important features of the original tumor, allowing, at the same time, experimental manipulation and in vivo amplification of the human cells. Here we present a detailed protocol for the generation of fluorescently labeled AML PDX models to monitor cell proliferation kinetics in vivo, at the single-cell level. Although experimental protocols for cell proliferation studies are well established and widespread, they are not easily applicable to in vivo contexts, and the analysis of related time-series data is often complex to achieve. To overcome these limitations, model-driven approaches can be exploited to investigate different aspects of cell population dynamics. Among the existing approaches, the ProCell framework is able to perform detailed and accurate stochastic simulations of cell proliferation, relying on flow cytometry data. In particular, by providing an initial and a target fluorescence histogram, ProCell automatically assesses the validity of any user-defined scenario of intra-tumor heterogeneity, that is, it is able to infer the proportion of various cell subpopulations (including quiescent cells) and the division interval of proliferating cells. Here we explain the protocol in detail, providing a description of our methodology for the conditional expression of H2B-GFP in human AML xenografts, data processing by flow cytometry, and the final elaboration in ProCell.
Original languageEnglish
Title of host publicationLeukemia Stem Cells
Subtitle of host publicationMethods and Protocols
EditorsCésar Cobaleda, Isidro Sánchez-García
PublisherSpringer
Pages241-258
Number of pages18
ISBN (Electronic)978-1-0716-0810-4
ISBN (Print)978-1-0716-0809-8
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2185
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Acute myeloid leukemia (AML)
  • Cell proliferation
  • Computational intelligence
  • Global optimization
  • Human xenografts
  • Mathematical modeling
  • Parameter estimation
  • Stochastic simulation

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