Citòmica Funcional

ICO - German Trias i Pujol

Josep Carreras Leukaemia Research Institute
Can Ruti Campus
Ctra de Can Ruti, Camí de les Escoles s/n
08916 Badalona, Barcelona, Spain

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Summary

The ability to bridge large-scale and single cell approaches at a functional level are key to identify biomarkers expressed in rare cells, and particularly in cancer stem cells (CSCs). We are an interdisciplinary group that investigates and develops key experimental approaches to understand the principles underlying the emergence and prevention of tumorigenesis and cancer therapy resistance, with the aim to translate biomedical science into integrated clinical practice and public health with certified transference processes in cooperation with trusted allies and partners.

Research

Since nearly three decades we study the implications of multidrug resistance transporters in stem and cancer cell biology. The cancer stem cell (CSC) model has been established as a cellular mechanism that contributes to phenotypic and functional heterogeneity in diverse cancer types. Recent observations, however, have highlighted many complexities and challenges: the CSC phenotype can vary substantially between patients, tumours may harbour multiple phenotypically or genetically distinct CSCs, metastatic CSCs can evolve from primary CSCs, and tumour cells may undergo reversible phenotypic changes.

Given that current clinical application of functional cytomics remains largely confined to few specific academic centres, our goal is to provide the patients with a wide range of scientific support strategies, through precision, oversight and accuracy.

Our current research

Our ongoing research projects cover innovative approaches to study the expression of primitive stem cell markers during origin, progression, maintenance of cancer and its management; the quality and safety assessment of haematopoietic blood progenitor and stem cell grafts; the role of myeloid derived suppressor cells in immunotherapy and targeted therapy for clinical decision-making; new cytomic strategies for whole blood and marrow immunostaining; the use of natural compounds for cancer treatment; and the accurate detection and significance of minimal residual disease in acute leukaemia.

Our goals

Our goal is to provide the patients with a wide range of scientific support strategies, through precision, oversight and accuracy to integrate:

• Clinical implementation of functional cytomic asays. Precision/personaized high-quality assays for individual patients, by integrating functional cytomics to accelerate new experimental approaches for ex vivo and in vivo drug sensitivity.

• Translation of functional screening into novel clinical strategies. Measure the impact of exogenous interventions such as drug exposure on tumour cell phenotype.

• The understanding of drug resistance and the prediction of effective drug combinations. 

• The reduction of costs by obtaining specialized instrumentation and personnel for execution of cytomic screens in partnership with stakeholders and biotechnological partners.

• Functional and immunophenotyping data sets aimed at understanding complex functional-to-phenotype correlations and, thereby, accelerate discovery of the biology of leukemogenesis as well as the clinical implementation of novel therapies.

Outcomes of our research

Because personalized medicine is able to provide the right therapy, in the right dose, improving patient stratification and prediction of disease, we expect to make available:

• Better diagnosis and earlier intervention. Cytome analysis could determine precisely whether patients are susceptible to drug toxicities, and choose the optimal treatment strategy.

• Individualized drug selection. Here we consider which molecular and functional models best predict how a patient will respond to a therapy to develop accurate and cost-effective tests. 

• Drug development challenges. A better understanding of clinical observations made during individualized drug development and conventional therapy will help to identify new disease subtypes and their associated molecular pathways, and design drugs that target them with more efficient trials. 

Collaborations

Clinical Cytometry Education Network (CCEN)

As a result of a generous grant from the Wallace H. Coulter Foundation (WHCF), the International Clinical Cytometry Society (ICCS) and the European Society for Clinical Cell Analysis (ESCCA) have partnered with the WHCF to form the Clinical Cytometry Education Network (CCEN) formerly called the Bi-Society Project. This is a multi-faceted collaboration endeavoring to create educational resources for the international clinical cytometry community.

Some of our goals include: to host practical clinical cytometry courses in both developed and developing countries, develop various e-learning content, including online courses, short video, and clinical cases, and to facilitate training of young professionals via the Visitor Training Program.

People

Selected publications

Petriz J

Flow cytometry of the side population (SP).

Curr Protoc Cytom 2013, Chapter 9 Unit9.23.
The side population (SP) has become an important hallmark for the definition of the stem-cell compartment, especially for the detection of stem cells and for their physical isolation by fluorescence-activated cell sorting (FACS). SP cells are CD34(-) and were discovered using ultraviolet excitation based on the efflux of Hoechst 33342 (Ho342). Although the method works as originally described, the protocol is difficult for most investigators to perform: first, because the ability to discriminate SP cells is based on the differential retention of Ho342 during a functional assay; second, because of the difficulties in setting the right experimental and acquisition conditions; and third, because analysis of the acquired data requires extensive expertise in flow cytometry to accurately detect the SP events. More recently, a new assay based on the efflux of Vybrant DyeCycle Violet stain (DCV) has been documented to discriminate SP cells. This unit contains many helpful pointers to aid the user in obtaining the best possible results with these assays.
Més informació
Balbuena J, Pachon G, Lopez-Torrents G, Aran JM, Castresana JS, Petriz J

ABCG2 is required to control the sonic hedgehog pathway in side population cells with stem-like properties.

Cytometry A Set 2011, 79 (9) 672-83. Epub 19 Jul 2011
The Sonic Hedgehog (Hh) pathway has been implicated in the maintenance of stem or progenitor cells in many adult tissues. Importantly, abnormal Hh pathway activation is also associated with initiation of neoplasia, but its role in tumor growth is still unclear. Here, we demonstrate that cyclopamine, a plant-derived alkaloid product used to inhibit the Hh signaling pathway, reduces the Side Population (SP) obtained by Hoechst 33342 (Ho342) dye measurements. In addition, cyclopamine is able to modulate, along with oxysterols and other products, the ABCG2 transporter by increasing Ho342 and mitoxantrone uptake. Therefore, if the SP is solely measured as a Ho342 dye extruding fraction, this may be significantly modulated by the inhibition of ABCG2 transport fraction, independently from the action of cyclopamine on the Hh pathway. Our results indicate that ABCG2 may act in the upstream regulation of the Hh signaling pathway to protect the stemness of the SP compartment, giving support to the cancer stem cell hypothesis and suggesting that ABCG2 is not only critical for increased resistance to anticancer agents.
Més informació
Cossarizza A et al

Guidelines for the use of flow cytometry and cell sorting in immunological studies.

Eur. J. Immunol. Oct 2017, 47 (10) 1584-1797. Més informació
Fornas O, Garcia J, Petriz J

Flow cytometry counting of CD34+ cells in whole blood.

Nat. Med. Jul 2000, 6 (7) 833-6. Més informació
Rico LG, Juncà J, Ward MD, Bradford JA, Bardina J, Petriz J

Acoustophoretic Orientation of Red Blood Cells for Diagnosis of Red Cell Health and Pathology.

Sci Rep 24 Oct 2018, 8 (1) 15705. Epub 24 Oct 2018
Distortions of the normal bi-concave disc shape for red blood cells (RBCs) appear in a number of pathologies resulting from defects in cell membrane skeletal architecture, erythrocyte ageing, and mechanical damage. We present here the potential of acoustic cytometry for developing new approaches to light-scattering based evaluation of red blood cell disorders and of the effects of storage and ageing on changes or damage to RBCs membranes. These approaches could be used to immediately evaluate the quality of erythrocytes prior to blood donation and following transfusion. They could also be applied to studying RBC health in diseases and other pathologies, such as artificial heart valve hemolysis, thermal damage or osmotic fragility. Abnormal distributions of erythrocytes can typically be detected after just 30 to 45 seconds of acquisition time using 1-2 µL starting blood volumes.
Més informació
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