Welcome back to another Preprint watch! This month, we want to highlight some exciting novel approaches in treating hematological malignancies, from improving cell therapies to unforeseen synergies between already discovered molecules. As always, if you have some new research preprints, send us your preprint here.
Cell therapy approaches
Mitofusin agonists enhances long-term engraftment and potency of HSC cultures in vivo
https://www.biorxiv.org/content/10.1101/2025.10.27.684807v1
Ex vivo treatment of cord blood HSCs with mitofusin agonists achieves over five-fold enhancement of long-term repopulating activity without expanding total cell numbers, through suppression of protein synthesis, increased autophagy, and lysosomal acidification. Mechanistically, fusion-competent MFN2 directly sequesters MTOR, promoting a catabolic state that preserves HSC potency. This novel MFN2-MTOR regulatory axis offers a promising strategy for expanding functional cord blood HSCs
Inhibition of farnesyltransferase activity diminishes hematopoietic stem cell ex vivo expansion ability
https://www.biorxiv.org/content/10.1101/2025.10.30.685582v1
From the authors: The rationale underlying this work is that elucidating the contribution of farnesyltransferase activity to hematopoietic stem cell expansion ex vivo will provide knowledge needed to better support hematopoietic stem cell expansion techniques for clinical applications that rely on this approach, like hematopoietic cell transplants and gene therapy. We discovered that pharmacological inhibition of farnesyltransferase activity with lonafarnib substantially diminished the ex vivo expansion potential of human and mouse hematopoietic stem cells, highlighting that hematopoietic stem cells rely on isoprenoids for their ex vivo maintenance.
2-Methoxyestradiol Treatment Prevents Graft-versus-Host Disease While Preserving Graft-versus-Leukemia Effect in Mice
https://www.biorxiv.org/content/10.1101/2025.08.11.667504v1
2ME2, a well-tolerated endogenous estrogen metabolite, significantly reduces GVHD mortality and morbidity in mice by suppressing donor CD4+ T cell accumulation and IFN-γ/IL-2 production, while leaving CD8+ cytolytic activity intact. Crucially, 2ME2 therapy preserves graft-versus-leukemia activity, addressing a key challenge in GVHD treatment.
Base editing of β0 thalassemia mutations as a therapeutic strategy for β-hemoglobinopathies: efficacy and genotoxicity studies
https://www.biorxiv.org/content/10.1101/2025.08.21.671453v1
From the authors: Preclinical safety and efficacy studies of a new gene therapy approach for patients with severe β-hemoglobinopathies.
Combinatorial base editing couples disease correction with lineage amplification in hematopoietic stem and progenitor cells
https://www.biorxiv.org/content/10.64898/2026.04.13.718029v1
A multiplex base editing strategy combining BCL11A enhancer disruption, HBG1/2 promoter editing, and introduction of a naturally occurring erythropoietin receptor truncation (tEPOR) synergistically enhances both HbF expression and erythroid proliferation beyond single-edit or Casgevy-treated controls in healthy, sickle cell, and β-thalassemia HSPCs. Edited HSPCs retain long-term engraftment and lineage repopulation capacity in vivo.
In-Depth Characterization of Stem Cell Potency and Genotoxicity for Clinical-Scale Ex Vivo CRISPR/Cas9 Gene Editing
https://www.biorxiv.org/content/10.64898/2026.01.08.698401v1
A comprehensive preclinical platform for ex vivo CRISPR/Cas9-AAV6 correction of Wiskott-Aldrich Syndrome in HSPCs achieves large-scale manufacturing while preserving stemness and multilineage function, with optimized AAV dosing and transient p53 modulation significantly improving long-term engraftment. Multi-tiered genotoxicity assessment reveals a favorable safety profile with minimal off-target risks, though longitudinal monitoring identifies rare donor-specific structural variants detectable only post-engraftment, underscoring the necessity of post-transplant surveillance.
CAR T cell therapy selectively depletes disease-driving mutant calreticulin cells in xenotransplants and human organoid models of myelofibrosis
https://www.biorxiv.org/content/10.64898/2026.01.28.702220v1
From the authors: A first-in-class CAR T-cell therapy targeting mutant calreticulin selectively depletes malignant stem cells in vivo and in fibrotic human organoids.
Differential TIM-3 glycosylation enables specific dual targeting CAR-T therapy in acute myeloid leukemia
https://www.biorxiv.org/content/10.64898/2026.04.22.719217v1
An IF-BETTER dual CAR approach, pairing CD33 and TIM-3 targeting, achieves potent antigen-restricted cytotoxicity against AML cell lines and primary blasts while sparing normal hematopoietic cells. Mechanistically, the TIM-3 scFv recognizes a glycosylation-dependent epitope, selectively enhanced by AML-specific hyper-fucosylation and hyper-sialylation.
Development of a Fully Non-Viral 1XX-enhanced BCMA CAR-T Cell Therapy for Multiple Myeloma
https://www.biorxiv.org/content/10.64898/2026.04.20.719660v1
From the authors: CRISPR-engineered, TRAC-targeted 1XX-BCMA CAR-T therapy with improved safety, potency, and persistence in relapsed and refractory multiple myeloma.
Novel small molecules and biologics
Discovery of A Small Molecule non-IMiD Degrader of ZBTB7A for the Treatment of β-hemoglobinopathies
https://www.biorxiv.org/content/10.1101/2025.09.17.676148v2
SH6 is a non-IMiD small molecule degrader of ZBTB7A that induces fetal hemoglobin in erythroid cells from healthy donors, sickle cell disease, and β-thalassemia patients in a CRBN-dependent manner, with efficacy confirmed in a humanized xenotransplantation model. SH6 outperforms currently available agents in vitro, shows synergy with hypomethylating agents, and displays a favorable toxicity profile.
A novel Fc-optimized antibody-drug conjugate targeting CD7 for the therapy of T-cell acute lymphoblastic leukemia
https://www.biorxiv.org/content/10.1101/2025.10.30.684608v1
From the authors: A novel antibody drug conjugate targeting CD7 showed efficient anti-leukemia activity in preclinical models of T-ALL.
CDK9 pharmacological inhibition with PRT2527 has antitumor activity in marginal zone lymphoma models and can improve the effects of BTK, PI3K, and BCL2 inhibitors
https://www.biorxiv.org/content/10.64898/2026.02.10.705084v1
From the authors: CDK9 inhibitor PRT2527 kills marginal zone lymphoma cells, regardless of whether they are resistant to other targeted drugs. PRT2527 boosts the effects of BTK, PI3K, and BCL2 inhibitors and alters immune-related gene expression.
CDK12/13 inhibitor, CTX-439, suppresses tumor growth and potentiates BCL-2 family blockade
https://www.biorxiv.org/content/10.64898/2026.02.20.706902v1
CTX-439, a novel oral CDK12/13 inhibitor, suppresses RNA Pol II S2 phosphorylation and downregulates DNA damage repair genes, exerting potent anti-cancer activity in vitro and in breast cancer PDX models. Mechanistically, CTX-439 reduces MCL-1 through transcriptional readthrough, shifting survival dependency to BCL-2 and BCL-xL, whose upregulation drives resistance.
Proteomic profiling reveals pleiotropic antimetabolite activity of triciribine in acute lymphoblastic leukemia
https://www.biorxiv.org/content/10.64898/2026.03.13.710465v1
Here, the authors examined triciribine, an Akt inhibitor that demonstrates potent cytotoxicity across diverse acute lymphoblastic leukemia (ALL) models. They report pleiotropic mechanisms — including impaired purine biosynthesis, DNA damage, translational stress, and CDK inhibition — rather than primary Akt suppression. Triciribine active metabolite TCN-P required adenosine kinase (ADK) for activity. ADK protein levels positively correlate with TCN sensitivity in both cell lines and primary patient samples, identifying ADK as a predictive biomarker for patient stratification.
New mechanisms, repurposing, and synergies
Pharmacological inhibition of LIN28A promotes imatinib sensitivity in CML resistance
https://www.biorxiv.org/content/10.64898/2026.04.27.720799v1
LIN28i-1632 synergizes potently with imatinib in resistant CML cells, reducing proliferation by 71% through proteomic remodeling: BCR-ABL downregulation, HMGA1 suppression, and AKT pathway attenuation, via RICTOR downregulation and PTEN restoration. Phosphoproteome and kinome profiling confirm suppressed AKT/RPS6K and CDK signaling alongside activation of apoptotic and G2/M checkpoint programs. These findings establish pharmacological LIN28 inhibition as a viable strategy to overcome BCR-ABL1-independent TKI resistance in CML.
Cholesterol Biosynthesis is a Targetable Vulnerability of CEBPA-mutant Acute Myeloid Leukemia
https://www.biorxiv.org/content/10.64898/2026.02.20.706425v1
From the authors: Induction of CEBPA-p30 by CRISPR/Cas gene editing in healthy human BM HSCs drives overt AML in vivo. TET2 and WT1 loss accelerate leukemogenesis, while GATA2 haploinsufficiency redirects differentiation toward erythroid precursors potentially driving acute erythroid leukemia. CEBPA-p30 AML exhibits cholesterol biosynthesis dependency, revealing a therapeutic vulnerability to statins.
Combined Menin and XPO1 inhibition drive synergistic antileukemic activity in KMT2Ar and NPM1-m AML
https://www.biorxiv.org/content/10.64898/2026.03.10.710924v1
Ziftomenib combined with the XPO1 inhibitor selinexor synergistically inhibits AML cell growth, suppresses colony formation in primary KMT2A-r progenitors, and induces apoptosis while sparing normal stem cells, with mechanistic studies revealing a novel role for XPO1 in stabilizing menin's chromatin binding and interactions with KMT2A. The combination downregulates HOXA9/MEIS1 and multiple menin-KMT2A target genes while promoting monocytic differentiation
Pitavastatin counteracts venetoclax resistance mechanisms in acute myeloid leukemia by depleting geranylgeranyl pyrophosphate
https://www.biorxiv.org/content/10.1101/2025.10.27.684888v1
Pitavastatin demonstrates potent cytotoxic activity in venetoclax-resistant AML cells harboring TP53 or FLT3 mutations, acting through p53-independent PUMA upregulation, MCL-1 downregulation, and suppression of mitochondrial oxidative metabolism. These pro-apoptotic effects depend on depletion of geranylgeranyl pyrophosphate (GGPP) and can be recapitulated by targeting GGPP synthesis or geranylgeranyltransferase-1.
A single phospho-LCK flow-cytometry readout predicts dasatinib sensitivity in paediatric T-cell acute lymphoblastic leukaemia
https://www.biorxiv.org/content/10.1101/2025.11.04.684929v1
Phospho-flow cytometry analysis of 28 pediatric T-ALL patient-derived xenografts identifies pLCK as the dominant single-marker predictor of dasatinib sensitivity, achieving excellent classification performance (AUC ≥0.9) with multivariate models adding minimal predictive value. These findings establish pLCK as a robust, scalable biomarker suitable for diagnostic integration.
Targeting MTHFD2 disrupts mitochondrial redox homeostasis and restores venetoclax sensitivity in acute myeloid leukemia
https://www.biorxiv.org/content/10.64898/2026.03.18.712743v1
MTHFD2 supports AML survival through mitochondrial one-carbon metabolism by sustaining de novo purine synthesis and NADH/NADPH production. Its loss increases mitochondrial superoxide and suppresses leukemic proliferation while sparing normal hematopoietic cells. The small-molecule inhibitor DS18561882 phenocopies genetic deletion, showing activity across 60 primary AML samples and synergizing with venetoclax in both treatment-naïve and resistant models.
Targeting WNK1 Releases Differentiation Block in Acute Myeloid Leukemia
https://www.biorxiv.org/content/10.64898/2026.04.22.720037v1
The authors identify WNK1 as a novel driver of AML differentiation arrest and a promising therapeutic target for expanding differentiation therapy to broader AML subtypes. WNK1 is overexpressed and active in AML patients, and its inhibition induces myeloid differentiation while suppressing growth, survival, and self-renewal in both cell lines and primary patient cells, with significant anti-tumor activity confirmed in vivo. Mechanistically, WNK1 inhibition derepress the MEK-ERK-C/EBPβ axis, upregulating myeloid differentiation genes.
DNA Demethylation is Dispensable for Venetoclax-HMA Synergy in Acute Myeloid Leukaemia
https://www.biorxiv.org/content/10.64898/2026.04.13.718134v1
The authors systematically compared the activities of hypomethylating agents with those of the DNMT1-selective inhibitor GSK5032, revealing that venetoclax synergy correlates with DNA damage-inducing properties rather than DNA demethylation. In fact, extensive demethylation alone fails to enhance — and may even antagonize — venetoclax activity. These findings reframe the mechanistic basis of VEN-HMA combinations,
5-Azacytidine incorporation into mRNAs disrupts translation and induces ribosome collisions
https://www.biorxiv.org/content/10.64898/2026.03.30.714548v1
The authors looked at the incorporation of 5-AzaC into mRNA. They observed C-to-G transversions at ribosomal A-sites, causing ribosome stalling and disome accumulation at C-rich codons within hours of treatment. This triggers both the integrated stress response and ribotoxic stress response in a GCN2- and ZAK-dependent manner, with loss of the RQC factor ZNF598 further sensitizing cells to 5-AzaC. These findings reveal translation disruption as an additional mechanism of 5-AzaC cytotoxicity alongside its established DNA hypomethylation effects.
Epigenetic control of S100A8/A9-driven monocytic inflammation licenses anti-leukemic functionality of immature NK cells during hematopoietic stem cell differentiation
https://www.biorxiv.org/content/10.64898/2026.03.25.714180v1
In this paper, the authors looked at healthy donor CD34+ stem cells. They realised that a subset of those HSPCs were epigenetically primed for S100A8/A9-driven inflammatory programs, promoting NK cell maturation and monocyte-NK crosstalk. 5-AzaC modulated these programs through upstream regulatory element rewiring. Moreover, they observed that S100A9 loss impairs myeloid differentiation and NK cell maturation via dysregulation of CEBPB, JUN, and NFIL3, while clinical re-analysis confirms that 5-AzaC responders display elevated S100A8/A9 alongside enhanced IFN-γ, IL6-JAK-STAT3, and TNF signaling. These findings position donor inflammatory myeloid programs as predictive biomarkers for 5-AzaC response and potential targets for enhancing graft-versus-leukemia activity post-transplant.
The Nicotinamide Salvage Pathway is a Metabolic Vulnerability of High-Risk MDS Stem Cells
https://www.biorxiv.org/content/10.64898/2025.12.19.695528v1
HR-MDS HSPCs exhibit elevated glycolytic, TCA cycle, and oxidative phosphorylation activity alongside increased mitochondrial complex I abundance, reflecting a heightened metabolic state compared to healthy HSPCs. NAMPT inhibition selectively depletes NAD(H), impairs oxidative capacity, reduces self-renewal and colony formation, and increases cell death specifically in HR-MDS HSPCs while sparing healthy cells.
Glutamine addiction is a therapeutic target to block emergency myelopoiesis
https://www.biorxiv.org/content/10.64898/2026.03.26.714544v1
From the authors: Olson et al. show that emergency myelopoiesis, the inflammatory overproduction of myeloid cells that drives regeneration, depends on Myc-driven mitochondrial respiration and glutamine addiction in hematopoietic progenitors. Targeting glutaminase in hematopoietic stem and progenitor cells suppresses pathological myelopoiesis, reduces tumor-promoting neutrophil production, and slows breast tumor growth.
Targeting glucocorticoid-induced CD20 activation in preclinical models of B-ALL
https://www.biorxiv.org/content/10.64898/2025.12.16.694550v1
Single-cell RNA sequencing of PDX models mimicking MRD identifies a transcriptionally distinct drug-tolerant population enriched for quiescence, inflammatory stress, and BCR pathway signatures, with CD20 consistently upregulated across diverse B-ALL subtypes. CD20 expression is induced by glucocorticoid exposure, and anti-CD20 monoclonal antibodies selectively eradicate MRD cells in vivo.
A dietary pan-amino acid dropout screen in vivo reveals a critical role for histidine in T-ALL
https://www.biorxiv.org/content/10.64898/2025.12.21.694897v1
A pan-amino acid dropout screen identifies histidine deprivation as uniquely antileukemic in NOTCH1-driven T-ALL, extending survival in a dose-dependent and well-tolerated manner. Mechanistically, histidine restriction triggers ribosome stalling, activating GCN2 to suppress cholesterol biosynthesis pathways critical for leukemic proliferation, an effect partially reversed by dietary cholesterol supplementation.
Thiol Scarcity in Cerebrospinal Fluid Renders Leptomeningeal Acute Lymphoblastic Leukaemia Therapeutically Vulnerable to Ferroptosis
https://www.biorxiv.org/content/10.64898/2025.12.15.693383v1
The CSF microenvironment's inherent thiol and cystine scarcity creates a pro-ferroptotic state in CNS-resident ALL cells, which become obligately dependent on LRP8-mediated selenium uptake and GPX4 activity for survival. Disrupting selenocysteine biosynthesis — genetically or via the FDA-approved agent Auranofin — induces synthetic lethality selectively in CNS-ALL models with excellent in vivo tolerability. These findings identify a mechanistically grounded, niche-exploiting therapeutic strategy with broad implications for leptomeningeal malignancies.
PTEN-loss confers dependence on the guanylate synthesis enzyme IMPDH in T-cell acute lymphoblastic leukemia
https://www.biorxiv.org/content/10.64898/2025.12.22.696045v1
From the authors: We find that the IMPDH inhibitors mycophenolic acid and mizoribine, which are currently used as well-tolerated immunosuppressants, exert anti-leukemic activity in an aggressive molecular subset of T-ALL cells that are associated with poor prognosis. The use of clinically approved compounds to exploit this vulnerability could lead to rapid drug repurposing.
Schlafen 11 (SLFN11) overexpression and nucleolar localization in response to bortezomib in multiple myeloma
https://www.biorxiv.org/content/10.64898/2026.02.06.704297v1
SLFN11 is consistently highly expressed in most multiple myeloma subtypes, correlating with super-enhancer-driven plasma cell transcriptional programs, and undergoes nucleolar translocation upon bortezomib treatment to suppress ribosomal RNA synthesis. SLFN11 knockout enhances bortezomib sensitivity while conferring exatecan resistance, establishing opposing roles in proteotoxic versus replication stress responses.
Expression profile of CASSIOPEIA patients refines prognostic value of MRD negativity in multiple myeloma
https://www.biorxiv.org/content/10.64898/2026.04.07.716874v1
Through RNA sequencing of 628 patients from the clinical trial CASSIOPEIA, the authors identified five transcriptomic subtypes. They stratified those subtypes into three risk categories, with 72-month Progression Free Survival (PFS) rates of 70%, 51%, and 27% for low, intermediate, and high-risk groups, respectively. Critically, MRD status fails to predict PFS or Overall Survival (OS) in the high-risk group, suggesting rapid expansion of aggressive minor clones despite daratumumab therapy.
Blog post contributed by Alessandro Donada, PhD (Bluesky: @alessandrodonada.bsky.social) of the ISEH Publications Committee.
Please note that the statements made by Simply Blood authors are their own views and not necessarily the views of ISEH. ISEH disclaims any or all liability arising from any author's statements or materials.