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2833 HSC/MPP-like Signature Predicts Inferior Outcome of Multiple Pediatric Leukemias

Program: Oral and Poster Abstracts
Session: 614. Acute Lymphoblastic Leukemias: Biomarkers, Molecular Markers, and Minimal Residual Disease in Diagnosis and Prognosis: Poster II
Hematology Disease Topics & Pathways:
Lymphoid Leukemias, ALL, Fundamental Science, Research, Translational Research, Diseases, Lymphoid Malignancies, Biological Processes, Pathogenesis
Sunday, December 8, 2024, 6:00 PM-8:00 PM

Jingliao Zhang1, Suyu Zong2*, Yongjuan Duan3*, Bei-bei Zhao4*, Jun Li5*, Xiaoli Chen6*, Wenyu Yang, MD7*, Peng Wu8*, Xiaofan Zhu, MD7 and Yingchi Zhang1*

1State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC, Tianjin, China
2State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Tianjin, China
3State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical Co, Tianjin, CHN
4State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical Colleg, Tianjin, China
5State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences&Peking Union Medical College, Tianjin, China
6State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology and Blood Diseases Hospital, CAMS & PUMC, Tianjin, CHN
7Department of Pediatric Hematology and Oncology, State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology& Blood Diseases Hospital, Chinese Academy of Medical Sciences& Peking Union Medical College, Tianjin, China
8State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Tianjin, China

Acute leukemia (AL) as the most common childhood malignancy widely encompasses acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), and Acute leukemias of ambiguous lineage (ALALs). The pathogenesis of AL is mainly attributed to the malignant transformation of hematopoietic stem/progenitor cells (HSPCs) into leukemic stem cells (LSCs) and the consequent blockage of normal hematopoietic differentiation. Currently, the mechanism of heterogeneous LSCs contributing to relapse and resistance to therapies remains to be challenging issues.

Following the designed sorting strategies to enrich leukemic cells and healthy bone marrow mononuclear cells (BMMCs) or stem/progenitor cells, we carried out high-throughput droplet-based 5′-single-cell RNA-sequencing (scRNA-seq) from four subtypes (B-ALL, T-ALL, AML, and MPAL) of pediatric leukemia at multiple stages and healthy donors, and identified manually curated pediatric hematopoietic differentiation landscape as HSC/MPP, erythroid, myeloid, and lymphoid) subclusters referring to expression patterns of marker genes. By projecting leukemic signatures from 23 pediatric acute leukemia patients at diagnosis onto this reference profiling of hematopoietic differentiation landscape. Notably, B-ALL and T-ALL blasts were dominantly projected onto LMPP-like and CLP-like subclusters, and the AML leukemic cells were broadly partitioned into granulocyte-monocyte progenitor-like and monocyte-like subclusters. Intriguingly, we observed significantly higher frequencies of HSC/MPP-like signatures in 4 of the 6 T-ALL, 2 of the 5 MPAL, and all of the 4 AML leukemic blasts, raising the lack of HSC/MPP signatures in our B-ALL cohort. With molecular heterogeneities in progenitor subclusters of MPALs, it was noteworthy that HSC/MPP-like signatures in T/Myeloid MPALs were presented with higher frequencies in contrast to those in B/Myeloid MPALs and the KMT2A-rearanged MPAL.

To understand the dynamics in proportional leukemic subclusters under chemotherapeutic pressure, we mapped transcriptomic signatures of longitudinal leukemia for the KMT2A-rearranged MPAL patient, who followed a standardized chemotherapy regimen for ALL and subsequently underwent umbilical cord blood transplantation due to progressive residual leukemia. Against a backdrop of continuous increases in measurable residual diseases (MRD) during hematopoiesis reconstitution, the LMPP-like subclusters demonstrated a decrease, while HSC/MPP-like subclusters strikingly emerged and gradually expanded over the course of treatment. Though not attaining dominance, the persistent growth of HSC/MPP-like signatures stood out as a salient observation within this dynamic context, inciting speculation of a potential correlation with chemoresistance.

Further assessment of differential expressed genes intersection within HSC/MPP subclusters across the three leukemia subtypes enriched 146 up-regulated genes and 122 down-regulated genes, with 24 genes exhibited ≥ 2-fold expression differences in the context of upregulated genes. We applied Cox proportional-hazards model to interrogate core biomarkers representing stemness that were also correlated with long-term outcomes across multiple pediatric leukemia, yielding to an ultimate 9 genes with robustness across leukemic subtypes. The predictive power of the 9-genes stemness-score in chemoresistance and long-term outcome was further validated from internal and external leukemic cohorts.

In this study, we utilized scRNA-seq to establish a differentiation landscape of normal HSPCs from HD and mapped multiple subtypes of leukemic cells on the hematopoietic differentiation roadmap. Notably, we found that HSC/MPP-like cells were commonly present in three out of the four leukemic subtypes. With longitudinal tracing, we discovered the gene expression signature of those leukemic cells was closely associated with leukemic molecular behaviors as well as clinical outcomes. Furthermore, the HSC/MPP-like signature was validated to be prominently correlated with chemoresistance and inferior long-term outcomes, warranting further mechanistic investigations.

Disclosures: No relevant conflicts of interest to declare.

*signifies non-member of ASH