Induced pluripotent stem cell models of myeloid malignancies and clonal evolution

Reilly, Andreea; Doulatov, Sergei

doi:10.1016/j.scr.2021.102195

Cited by 5 publications

(5 citation statements)

References 110 publications

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“…Here, we reported detailed comparative analyses of HPCs that were differentiated from RALD patient-derived iPSCs. Compared with iPSC-based modeling of hematologic malignancies that typically contain multiple mutated oncogenes [ 33 , 41 , 42 ], our RALD-iPSCs acted as an invaluable tool to investigate the considerable effect of KRAS (G13C) on cellular functions within an isogenic HPC pair. Besides studies examining the impact of heterozygous mutation of NRAS (G12D) [ 43 ] or KRAS (G12D) [ 44 , 45 ] for murine HPCs, to the best of our knowledge, this is the first study to analyze the effects of an oncogenic RAS protein expressed under endogenous control in the context of non-immortalized, human diploid HPCs.…”

Section: Discussionmentioning

confidence: 99%

Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells

Lin,

Takagi,

Kubara

et al. 2024

Stem Cell Res Ther

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Background Although oncogenic RAS mutants are thought to exert mutagenic effects upon blood cells, it remains uncertain how a single oncogenic RAS impacts non-transformed multipotent hematopoietic stem or progenitor cells (HPCs). Such potential pre-malignant status may characterize HPCs in patients with RAS-associated autoimmune lymphoproliferative syndrome-like disease (RALD). This study sought to elucidate the biological and molecular alterations in human HPCs carrying monoallelic mutant KRAS (G13C) with no other oncogene mutations. Methods We utilized induced pluripotent stem cells (iPSCs) derived from two unrelated RALD patients. Isogenic HPC pairs harboring either wild-type KRAS or monoallelic KRAS (G13C) alone obtained following differentiation enabled reliable comparative analyses. The compound screening was conducted with an established platform using KRAS (G13C) iPSCs and differentiated HPCs. Results Cell culture assays revealed that monoallelic KRAS (G13C) impacted both myeloid differentiation and expansion characteristics of iPSC-derived HPCs. Comprehensive RNA-sequencing analysis depicted close clustering of HPC samples within the isogenic group, warranting that comparative studies should be performed within the same genetic background. When compared with no stimulation, iPSC-derived KRAS (G13C)-HPCs showed marked similarity with the wild-type isogenic control in transcriptomic profiles. After stimulation with cytokines, however, KRAS (G13C)-HPCs exhibited obvious aberrant cell-cycle and apoptosis responses, compatible with "dysregulated expansion," demonstrated by molecular and biological assessment. Increased BCL-xL expression was identified amongst other molecular changes unique to mutant HPCs. With screening platforms established for therapeutic intervention, we observed selective activity against KRAS (G13C)-HPC expansion in several candidate compounds, most notably in a MEK- and a BCL-2/BCL-xL-inhibitor. These two compounds demonstrated selective inhibitory effects on KRAS (G13C)-HPCs even with primary patient samples when combined. Conclusions Our findings indicate that a monoallelic oncogenic KRAS can confer dysregulated expansion characteristics to non-transformed HPCs, which may constitute a pathological condition in RALD hematopoiesis. The use of iPSC-based screening platforms will lead to discovering treatments that enable selective inhibition of RAS-mutated HPC clones.

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Section: Discussionmentioning

confidence: 99%

Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells

Lin,

Takagi,

Kubara

et al. 2024

Stem Cell Res Ther

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“…Sequence of phenotype changes associated to mutations reveals that in the first stage, mutations may increase self-renewal, but in the following stages, another driver mutation may help the HSCs to become a leukemic stem cancer cell. This process has been shown also to develop in male germ cells, favoring clonal expansion and the transmission of mutations (Jan and Majeti 2013;Minussi et al 2021;Reilly and Doulatov 2021).…”

Section: Aging and Genomic Instability In Stem Cellsmentioning

confidence: 99%

A Multilevel Approach to the Causes of Genetic Instability in Stem Cells

González

2022

Handbook of Stem Cell Therapy

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This chapter addresses the genetic instability in stem cells, a central feature that is an important determinant for the safety and effectiveness of cell-based therapies. First, DNA damage response mechanism and the gene network that regulates the DNA integrity homeostasis are revisited, focusing on relationship between genetic integrity and stemness maintenance. Also, several factors have been included that influence genetic instability in vivo, i.e., ROS generation and inflammation, and in vitro regarding cell isolation, culture conditions, i.e., oxygen levels and the use of feeder layers and different carriers, splitting procedures, passage number, etc. Telomere shortening, replicative senescence aneuploidy, and the senescence-associated secretory phenotype are different processes involved in deterioration of stem cells abilities for homing, reparability, and paracrine modulation. Moreover, less effective DNA repair upon senescence increases the propensity of developing tumors for stem cells that is one the main concerns related to genetic instability in stem cells. Nuclear organization and their determinants linked to chromosome aberrations and aneuploidy in stem cells and those epigenetic changes affecting stability are addressed. Differences between adipose, embryonic, and induced pluripotent stem cells are analyzed regarding to their ontogeny, changes in culture, and variation in proliferative capacity and stemness. The effect of reprogramming methods in genetic instability and variation in mutagenicity according to genes utilized for pluripotency induction, i.e., Yamanaka's factors and others, is discussed. Donor-related factors, i.e., age, smoking, and alcohol consumption, comorbidities, i.e., obesity, insulin resistance, diabetes, are mentioned for wide evaluation of genetic instability. The next years must witness consensus protocols that will contribute to control the effects of factors that alter stem cell genetic stability to increase the security and applicability of cell-based therapy.

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“…Direct conversion approaches to generate cells of the hematopoietic lineage from fibroblasts using a one-step approach or directed differentiation from human embryonic stem cells have been used as an alternative to somatic cell reprogramming followed by hematopoietic differentiation of iPSCs [51]. The use of iPSC model for hematological disorders with a focus on patient-specific iPSCs has been reviewed [50,[52][53][54]. The generation of isogenic pairs of normal and mutated iPSC lines using gene editing methodology helps in understanding the key role of specific mutations.…”

Section: Transposon-based Insertional Mutagenesismentioning

confidence: 99%

“…The current review focuses on the development of iPSC models for understanding initiation and progression of hematological malignancies with a primary focus on using genetically engineered iPSCs as an alternative to patient-derived iPSCs. While the importance of hematopoietic differentiation of iPSC in disease modeling has been reviewed elsewhere [53,54], the present review highlights the development of 3-dimensional culture protocols for in vitro differentiation of iPSCs towards hematopoietic lineage.…”

Section: Transposon-based Insertional Mutagenesismentioning

confidence: 99%

Harnessing the Power of Induced Pluripotent Stem Cells and Gene Editing Technology: Therapeutic Implications in Hematological Malignancies

Sidhu

Barwe

Pillai

et al. 2021

Cells

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In vitro modeling of hematological malignancies not only provides insights into the influence of genetic aberrations on cellular and molecular mechanisms involved in disease progression but also aids development and evaluation of therapeutic agents. Owing to their self-renewal and differentiation capacity, induced pluripotent stem cells (iPSCs) have emerged as a potential source of short in supply disease-specific human cells of the hematopoietic lineage. Patient-derived iPSCs can recapitulate the disease severity and spectrum of prognosis dictated by the genetic variation among patients and can be used for drug screening and studying clonal evolution. However, this approach lacks the ability to model the early phases of the disease leading to cancer. The advent of genetic editing technology has promoted the generation of precise isogenic iPSC disease models to address questions regarding the underlying genetic mechanism of disease initiation and progression. In this review, we discuss the use of iPSC disease modeling in hematological diseases, where there is lack of patient sample availability and/or difficulty of engraftment to generate animal models. Furthermore, we describe the power of combining iPSC and precise gene editing to elucidate the underlying mechanism of initiation and progression of various hematological malignancies. Finally, we discuss the power of iPSC disease modeling in developing and testing novel therapies in a high throughput setting.

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Induced pluripotent stem cell models of myeloid malignancies and clonal evolution

Cited by 5 publications

References 110 publications

Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells

Monoallelic KRAS (G13C) mutation triggers dysregulated expansion in induced pluripotent stem cell-derived hematopoietic progenitor cells

A Multilevel Approach to the Causes of Genetic Instability in Stem Cells

Harnessing the Power of Induced Pluripotent Stem Cells and Gene Editing Technology: Therapeutic Implications in Hematological Malignancies

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