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Klusmann, J.-H.

Publications and source records attributed to Klusmann, J.-H..

11 recordsLinked to original sources

Targeting IGF2BP1 Creates a Mitotic Vulnerability in Colorectal Carcinoma

Background Mitotic dysregulation is a common feature of colorectal cancer (CRC), yet the contribution of post-transcriptional mechanisms remain incompletely understood. We investigated how the oncofetal RNA-binding protein IGF2BP1 supports mitotic integrity and exposes therapeutic vulnerabilities in CRC. Methods We integrated patient cohort analyses with genetic and pharmacological perturbation of IGF2BP1, transcriptomics, proteomics, RNA-binding assays and functional studies. Xenografts assessed tumor formation, while BTYNB-paclitaxel responses were examined in CRC cell lines and patient-derived organoids. Results IGF2BP1 expression was elevated in CRC, increased with tumor stage, and associated with poor survival and proliferative gene signatures. Genetic depletion or pharmacological inhibition of IGF2BP1 impaired cell viability, induced mitotic abnormalities, and promoted apoptosis, whereas its re-expression enhanced tumor formation in vivo. Integrated molecular analyses identified the chromosomal passenger complex (CPC) as a previously unrecognized downstream target of IGF2BP1. IGF2BP1 bound transcripts encoding CPC components and its depletion consistently reduced their expression across multiple cancer cell models. BTYNB disrupted IGF2BP1 association with INCENP, CDCA8, and BIRC5 transcripts and accelerated INCENP mRNA decay. Functionally, INCENP depletion recapitulated the mitotic defects and impaired viability observed upon IGF2BP1 inhibition. Exploiting this mitotic vulnerability, BTYNB synergized with paclitaxel in CRC cell lines and across four independent IGF2BP1-positive patient-derived organoid models. Conclusions Our findings uncover an IGF2BP1-dependent post-transcriptional regulatory mechanism sustaining CPC expression and mitotic integrity in CRC. Targeting this mechanism exposes a therapeutically exploitable mitotic vulnerability and provides a rationale for combining IGF2BP1 inhibition with microtubule-targeting agents.

cancer biology↗

Integrative enhancer discovery identifies functional enhancer dependencies in pediatric acute myeloid leukemia

Pediatric acute myeloid leukemia (AML) is driven by aberrant transcriptional programs sustained by poorly defined cis-regulatory mechanisms. To systematically identify functional enhancer dependencies, we developed an integrative enhancer discovery strategy that combines H3K27ac CUT&Tag profiling, enhancer-associated transcription, and CRISPR interference (CRISPRi) screening. By leveraging enhancer-associated transcription to prioritize candidate regulatory elements, we identified 321 leukemia-associated enhancers for functional interrogation. This approach uncovered the hematopoietic MYB enhancer (H-ME) within the HBS1L-MYB-AHI1 locus as a critical regulator of leukemic growth. H-ME repression reduced chromatin accessibility and active histone marks at the MYB promoter, suppressed MYB expression, and induced differentiation-associated transcriptional programs. In contrast, selective depletion of the enhancer-associated transcript had no effect on MYB expression or leukemic proliferation, demonstrating that enhancer activity resides within the underlying regulatory DNA element rather than its mature RNA product. H-ME exhibited preferential activity in megakaryocytic leukemia, and its perturbation impaired leukemic growth in primary patient-derived models in vitro and in vivo. Together, our findings establish an integrative framework for the systematic discovery of functional enhancer dependencies and identified H-ME as an RNA-independent regulator of MYB in pediatric AML.

cancer biology↗

Lineage-restricted dependency on an oncofetal SNHG29-IGF2BP1 RNA axis in acute megakaryoblastic leukemia

Acute megakaryoblastic leukemia (AMKL) is a rare, aggressive subtype of acute myeloid leukemia with developmental origins in early childhood. To uncover long noncoding RNAs (lncRNAs) sustaining this high-risk malignancy, we conducted CRISPR interference screens targeting lncRNAs overexpressed in primary AMKL samples. This analysis identified SNHG29 as a previously unrecognized lineage-specific dependency, whose silencing profoundly impaired leukemic proliferation and clonogenic growth in vitro and reduced leukemic burden in vivo. RNA pulldown coupled with proteomic analysis revealed that SNHG29 interacts with the oncofetal RNA-binding protein IGF2BP1, which is aberrantly expressed in AMKL. SNHG29 was required to maintain expression of IGF2BP1 target transcripts, including MYC- and E2F-driven proliferative programs, thereby reinforcing fetal transcriptional programs essential for leukemic maintenance. Pharmacologic inhibition of IGF2BP1-RNA interactions with the small-molecule BTYNB induced potent and selective cytotoxicity in patient-derived AMKL models. Together, these findings uncover a developmental co-dependency between SNHG29 and IGF2BP1 that defines a lineage-restricted oncogenic circuit and an actionable therapeutic vulnerability in AMKL.

cancer biology↗

The ITCC-P4 PDX platform of pediatric cancers for preclinical testing

Cancer is the leading cause of disease-related deaths among children in high-income countries. Tumor heterogeneity and lack of mechanism-of-action-based therapeutic options are key challenges to overcome in order to improve pediatric cancer patients survival. Here, we report the EU-IMI-2 funded public-private partnership "ITCC-Pediatric Preclinical Proof-of-Concept Platform" (ITCC-P4), which has built a large repertoire of patient-derived xenograft (PDX) models, representing all major solid pediatric cancer types, for in vivo drug testing. Three-hundred-fifty-three PDX models from diagnostic and relapsed pediatric cancers have been established and molecularly characterized, together with matched germline/tumor samples. As proof-of-concept, we present in vivo drug screening data in neuroblastoma and rhabdomyosarcoma models. PDX data, accessible at http://r2platform.com/itcc-p4, allow the selection of models based on oncogenic drivers and/or potential biomarkers for preclinical testing. Operated by a non-profit entity (www.itccp4.com), this sustainable platform aids academic and industrial researchers in developing and prioritizing innovative therapies for pediatric cancer. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/703023v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@195ba30org.highwire.dtl.DTLVardef@f2c2d9org.highwire.dtl.DTLVardef@1d63f4dorg.highwire.dtl.DTLVardef@d60027_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

A platform of robust patient-derived leukemia models covering subgroups for which no cell lines exist

Preclinical cancer research requires robust model systems, especially for poor prognosis entities like acute myeloid leukemia (AML), a highly aggressive blood cancer. Here, primary tumor cells from 137 AML patients of all age groups were transplanted into immune compromised mice to generate patient-derived xenografts (PDX). From these, 23 models enable robust, virtually endless serial re-transplantation and are amenable to lentiviral genetic engineering (*PDX AML models). These models primarily originate from patients with highly aggressive, relapsed disease. Comprehensive genomic, transcriptomic, and epigenomic analyses confirmed that they replicate primary AML biology more faithfully than conventional cell lines. Notably, *PDX AML models include AML subgroups that are underrepresented or absent in existing model systems, such as cytogenetically normal or IDH1/2-mutant AML. They withstand freeze-thaw cycles, making them suitable for broad distribution and reproducibility across research institutions. Luciferase-based in vivo imaging enables real-time monitoring of tumor progression and treatment responses in preclinical trials. Surprisingly, long-term treatment, including repeated cytarabine therapy over a period of one year, showed a gradual reduction in leukemia cell proliferation, which decreased continuously after each treatment block. Collectively, our *PDX models represent a robust, versatile, and relevant platform that holds great promise to accelerate translational research for the benefit of cancer patients. Visual Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/677299v1_ufig1.gif" ALT="Figure 1"> View larger version (23K): org.highwire.dtl.DTLVardef@1ead31eorg.highwire.dtl.DTLVardef@1ca702corg.highwire.dtl.DTLVardef@18a7137org.highwire.dtl.DTLVardef@edbe6_HPS_FORMAT_FIGEXP M_FIG C_FIG Key PointsO_LIWe present new robust AML PDX models covering subgroups for which no cell lines exist for use in various ex vivo and in vivo applications. C_LIO_LI*PDX models enable serial transplantation, genetic engineering and better representation of primary AML biology than cell lines. C_LIO_LIOne-year in vivo trials mimicking clinical chemotherapy showed surprising gradual decline in leukemia growth after each treatment block. C_LI

cancer biology↗

The activation of TP53 pathway is a therapeutic vulnerability in NUP98::KDM5A+ pediatric AML

NUP98::KDM5A-rearranged pediatric acute myeloid leukemia (AML) is a rare, infancy-predominant entity with dismal outcome and no targeted therapeutic options. Given its suspected fetal origin, we hypothesized that leukemic maintenance depends on developmentally restricted vulnerabilities embedded within fetal hematopoietic programs. Using matched fetal and adult hematopoietic stem and progenitor cell models, we integrated transcriptomic and proteomic profiling with CRISPR-Cas9 screenings to define ontogeny-specific dependencies in NUP98::KDM5A leukemia. Fetal-derived NUP98::KDM5A leukemias exhibited greater in vivo aggressiveness, and retained fetal transcriptional signatures compared with adult counterparts. A comparative CRISPR-Cas9 screen using a library targeting fetal gene programs, conducted in both fetal- and adult-derived NUP98::KDM5A leukemias, identified the AAA ATPase TRIP13 as a selective and essential dependency in the fetal context. Mechanistically, TRIP13 physically interacted with the TP53 phosphatase PPM1D/WIP1, resulting in suppression of TP53 activation. Genetic ablation of Trip13, or pharmacologic inhibition using DCZ0415, a small-molecule inhibitor targeting TRIP13, reactivated TP53, induced G2/M arrest and mitochondrial apoptosis, and depleted leukemic cells in vitro; these effects were fully rescued by TP53 loss. In competitive transplantation assays, Trip13 ablation significantly impaired leukemic fitness in vivo. Together, these findings define a developmentally restricted TRIP13-PPM1D-TP53 survival axis in NUP98::KDM5A AML and provide mechanistic proof-of-concept that reactivation of the TP53 signaling pathway via TRIP13 inhibition represents a therapeutically targetable vulnerability in this high-risk pediatric leukemia. Visual abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=121 SRC="FIGDIR/small/675580v4_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@143d103org.highwire.dtl.DTLVardef@14f3225org.highwire.dtl.DTLVardef@655541org.highwire.dtl.DTLVardef@c021ae_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Single cell transcriptional evolution of myeloid leukaemia of Down syndrome

Children with Down syndrome have a 150-fold increased risk of developing myeloid leukaemia (ML-DS). Unusually for a childhood leukaemia, ML-DS arises from a preleukaemic state, termed transient abnormal myelopoiesis (TAM), via a conserved sequence of mutations. Here, we examined the relationship between the genetic and transcriptional evolution of ML-DS from natural variation; a rich collection of primary patient samples and fetal tissues with a range of constitutional karyotypes. We distilled transcriptional consequences of each genetic step in ML-DS evolution, utilising single cell mRNA sequencing, complemented by phylogenetic analyses in progressive disease. We found that transcriptional changes induced by the TAM-defining GATA1 mutations are retained in, and account for most of the ML-DS transcriptome. The GATA1 transcriptome pervaded all stages of ML-DS, including progressive disease that had undergone genetic evolution. Our approach delineates the transcriptional evolution of ML-DS and provides an analytical blueprint for distilling consequences of mutations within their pathophysiological context.

cancer biology↗

RUNX1 isoform disequilibrium in the development of trisomy 21 associated myeloid leukemia

Aneuploidy is a hallmark of cancer, but its complex nature limits our understanding of how it drives oncogenesis. Gain of chromosome 21 (Hsa21) is among the most frequent aneuploidies in leukemia and is associated with markedly increased leukemia risk. Here, we propose that disequilibrium of the RUNX1 isoforms is key to the pathogenesis of trisomy 21 (i.e. Down syndrome)-associated myeloid leukemia (ML-DS). Hsa21-focused CRISPR-Cas9 screens uncovered a strong and specific RUNX1 dependency in ML-DS. Mechanistic studies revealed that excess of RUNX1A isoform - as seen in ML-DS patients - synergized with the pathognomonic Gata1s mutation in leukemogenesis by displacing RUNX1C from its endogenous binding sites and inducing oncogenic programs in complex with the MYC cofactor MAX. These effects were reversed by restoring the RUNX1A:RUNX1C equilibrium or pharmacological interference with MYC:MAX dimerization. Our study highlights the importance of alternative splicing in leukemogenesis, even on a background of aneuploidies, and opens new avenues for developing specific and targeted therapies. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/483334v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@16c5a95org.highwire.dtl.DTLVardef@af0ec7org.highwire.dtl.DTLVardef@f36bcborg.highwire.dtl.DTLVardef@129e832_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

Combining LSD1 and JAK-STAT inhibition targets Down syndrome-associated myeloid leukemia at its core

Children with Down syndrome (DS) are predisposed to developing megakaryoblastic leukemia (ML-DS) and often experience severe toxicities from chemotherapy, highlighting the need for targeted therapies with beneficial risk profiles. The genomic landscape of ML-DS is characterized by a combination of mutations in signaling pathway genes and epigenetic modifiers, while aberrant lysine specific demethylase 1 (LSD1) and JAK-STAT activation have both been implicated in leukemogenesis. Here, we demonstrate that combined LSD1 and JAK1/2 inhibition exerts synergistic anti-leukemic effects specifically in ML-DS, both in vitro and in patient derived xenografts in vivo. The JAK1/2 inhibitor ruxolitinib enhanced the LSD1 inhibitor-induced differentiation, proliferation arrest and apoptosis in patient-derived leukemic blasts. At the transcriptional level, the combination synergistically repressed gene expression signatures essential for cell division. We further observed an immunogenic gene expression pattern in the form of increased cytokine signaling, which - by sensitizing ML-DS blasts to the JAK-STAT signaling blockade induced by ruxolitinib - could explain the increased susceptibility of ML-DS blasts to combination therapy. Taken together, we establish combined LSD1 and JAK-STAT inhibition as an efficacious therapeutic regimen specifically designed to target important steps in ML-DS leukemogenesis, paving the way for targeted therapies in this entity.

cancer biology↗

Naked mole-rat hematopoietic stem and progenitors are highly quiescent with an inherent myeloid bias

Naked mole-rats (NMRs) are the longest-lived rodents yet their stem cell characteristics remain enigmatic. Here we comprehensively mapped the NMR hematopoietic landscape and identified unique features likely contributing to longevity. Adult NMRs form red blood cells in spleen and marrow, which is a neotenic trait. A myeloid bias towards granulopoiesis in concert with decreased B-lymphopoiesis defines the marrow composition, resembling fetal leukopoiesis. Very similar to primates, the primitive stem cell compartment is marked by CD34 and THY1. Remarkably, stem and progenitor respiration rates are as low as in human cells, while NMR cells show a strong expression signature for fatty acid metabolism. The pool of quiescent stem cells is higher than in mice, and the cell cycle of hematopoietic cells is prolonged. Our work provides a platform to study immunology and stem cell biology in an animal model of exceptional longevity. TeaserJuvenile features of hematopoiesis shape the blood system of the longest-lived rodent.

cell biology↗

The megakaryocytic transcription factor ARID3A suppresses leukemia pathogenesis

Given the plasticity of hematopoietic stem/progenitor cells, multiple routes of differentiation must be blocked during acute myeloid leukemia pathogenesis - the molecular basis of which is incompletely understood. Here we report that post-transcriptional repression of transcription factor ARID3A by miR-125b is a key event in megakaryoblastic leukemia (AMKL) pathogenesis. AMKL is frequently associated with trisomy 21 and GATA1 mutations (GATA1s), and children with Down syndrome are at a high risk of developing this disease. We show that chromosome 21-encoded miR-125b synergizes with Gata1s to drive leukemogenesis in this context. Leveraging forward and reverse genetics, we uncover Arid3a as the main miR-125b target underlying this synergy. We demonstrate that during normal hematopoiesis this transcription factor promotes megakaryocytic differentiation in concert with GATA1 and mediates TGF{beta}-induced apoptosis and cell cycle arrest in complex with SMAD2/3. While Gata1s mutations perturb erythroid differentiation and induce hyperproliferation of megakaryocytic progenitors, intact ARID3A expression assures their megakaryocytic differentiation and growth restriction. Upon knockdown, these tumor suppressive functions are revoked, causing a dual megakaryocytic/erythroid differentiation blockade and subsequently AMKL. Inversely, restoring ARID3A expression relieves the megakaryocytic differentiation arrest in AMKL patient-derived xenografts. This work illustrates how mutations in lineage-determining transcription factors and perturbation of post-transcriptional gene regulation interplay to block multiple routes of hematopoietic differentiation and cause leukemia. Surmounting this differentiation blockade in megakaryoblastic leukemia by restoring the tumor suppressor ARID3A represents a promising strategy for treating this lethal pediatric disease. Key pointsO_LIRepression of megakaryocytic transcription factor ARID3A by miR-125b synergizes with GATA1s to induce leukemia C_LIO_LIRestoring ARID3A expression relieves megakaryocytic differentiation arrest in megakaryoblastic leukemia C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=198 SRC="FIGDIR/small/440795v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@146c2a7org.highwire.dtl.DTLVardef@9599fdorg.highwire.dtl.DTLVardef@1b0d22borg.highwire.dtl.DTLVardef@1b61447_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗