Acute myeloid leukemia (AML) is an often devastating hematologic malignancy with 5-year overall survival lingering near 20%. Acquiring a deeper understanding of molecular underpinnings of leukemogenesis will provide a basis for developing more effective therapeutic strategies for patients with AML.
Here, we identified overexpression of hnRNP K as a recurrent abnormality in a subset (~20%) of AML patients. High levels of this RNA-binding protein associated with inferior clinical outcomes in de novo AML. Thus, to evaluate its putative oncogenic capacity in myeloid disease, we overexpressed hnRNP K in murine hematopoietic stem and progenitor cells isolated from fetal liver cells (FLCs). We revealed that hnRNP K-overexpression alters self-renewal capacity and differentiation potential of these cells in vitro. Such findings were recapitulated in vivo, as murine recipients of hnRNP K-overexpressing FLCs developed fatal myeloproliferative phenotypes.
To elucidate mechanisms by which hnRNP K overexpression causes myeloid neoplasia, we took an unbiased approach utilizing RNA-immunoprecipitation sequencing (fRIP-Seq). Among RNA transcripts interacting with hnRNP K was RUNX1—a pivotal transcriptional regulator of definitive hematopoiesis commonly mutated or translocated in AML. Consensus hnRNP K binding sites were identified in the 5’ UTR and near the 3’ splice site in intron 5-6 of RUNX1. Fluorescence anisotropy studies confirmed these interactions were direct, and abrogated when hnRNP K binding sites within RUNX1 were mutated. Manipulating hnRNP K expression in human cell lines and murine FLCs substantially altered RUNX1 splicing surrounding exon 6. RNA-sequencing of FLCs confirmed these data, exposing RUNX1 as a significantly differentially spliced entity in the context of hnRNP K overexpression. Importantly, the protein product of this spliced product (RUNX1ΔEx6) exhibited disparate transcriptional activity in reporter assays compared to full-length RUNX1. Furthermore, we identified KH3 as the hnRNP K domain most critical for these splicing alterations; deletion of KH3 markedly abrogated hnRNP K overexpression phenotypes in vitro.
In sum, we established hnRNP K as an oncogene in myeloid leukemia that binds RUNX1 RNA, altering its splicing and subsequent transcriptional activity. These findings shed light on a mechanism of myeloid leukemogenesis, paving the way for drug discovery efforts to improve outcomes for patients with this disease.
","abstract_html":"<p>Acute myeloid leukemia (AML) is an often devastating hematologic malignancy with 5-year overall survival lingering near 20%. Acquiring a deeper understanding of molecular underpinnings of leukemogenesis will provide a basis for developing more effective therapeutic strategies for patients with AML.</p> <p>Here, we identified overexpression of hnRNP K as a recurrent abnormality in a subset (~20%) of AML patients. High levels of this RNA-binding protein associated with inferior clinical outcomes in <em>de novo </em>AML. Thus, to evaluate its putative oncogenic capacity in myeloid disease, we overexpressed hnRNP K in murine hematopoietic stem and progenitor cells isolated from fetal liver cells (FLCs). We revealed that hnRNP K-overexpression alters self-renewal capacity and differentiation potential of these cells <em>in vitro. </em>Such findings were recapitulated <em>in vivo, </em>as murine recipients of hnRNP K-overexpressing FLCs developed fatal myeloproliferative phenotypes.</p> <p>To elucidate mechanisms by which hnRNP K overexpression causes myeloid neoplasia, we took an unbiased approach utilizing RNA-immunoprecipitation sequencing (fRIP-Seq). Among RNA transcripts interacting with hnRNP K was <em>RUNX1</em>—a pivotal transcriptional regulator of definitive hematopoiesis commonly mutated or translocated in AML. Consensus hnRNP K binding sites were identified in the 5’ UTR and near the 3’ splice site in intron 5-6 of <em>RUNX1. </em>Fluorescence anisotropy studies confirmed these interactions were direct, and abrogated when hnRNP K binding sites within <em>RUNX1 </em>were mutated. Manipulating hnRNP K expression in human cell lines and murine FLCs substantially altered <em>RUNX1 </em>splicing surrounding exon 6. RNA-sequencing of FLCs confirmed these data, exposing <em>RUNX1 </em>as a significantly differentially spliced entity in the context of hnRNP K overexpression. Importantly, the protein product of this spliced product (RUNX1ΔEx6) exhibited disparate transcriptional activity in reporter assays compared to full-length RUNX1. Furthermore, we identified KH3 as the hnRNP K domain most critical for these splicing alterations; deletion of KH3 markedly abrogated hnRNP K overexpression phenotypes <em>in vitro. </em></p> <p>In sum, we established hnRNP K as an oncogene in myeloid leukemia that binds <em>RUNX1 </em>RNA, altering its splicing and subsequent transcriptional activity. These findings shed light on a mechanism of myeloid leukemogenesis, paving the way for drug discovery efforts to improve outcomes for patients with this disease.</p>","abstract_has_math":false,"creators":["Aitken, Marisa","https://orcid.org/0000-0002-7937-531X
"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation (PhD)","degree_discipline":null,"degree_department":null,"school":null,"contributors":["Sean Post, Ph.D.","Scott Evans, M.D.","Marina Konopleva, M.D., Ph.D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-05-01T07:00:00Z","date_published":"2020-05-01T07:00:00Z","updated_at":"2026-07-24T05:50:31Z","subjects":["hnRNP K","RNA binding proteins","RNA splicing","AML","myeloproliferative disease","mouse model","Animal Experimentation and Research","Cancer Biology","Cell Biology","Genetics","Hemic and Lymphatic Diseases","Immune System Diseases","Immunopathology","Medicine and Health Sciences"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://digitalcommons.library.tmc.edu/utgsbs_dissertations/994","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Sean Post, Ph.D.","Scott Evans, M.D.","Marina Konopleva, M.D., Ph.D."]},{"key":"dc:creator","label":"Author","values":["Aitken, Marisa","https://orcid.org/0000-0002-7937-531X
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Here, we identified overexpression of hnRNP K as a recurrent abnormality in a subset (~20%) of AML patients. High levels of this RNA-binding protein associated with inferior clinical outcomes in de novo AML. Thus, to evaluate its putative oncogenic capacity in myeloid disease, we overexpressed hnRNP K in murine hematopoietic stem and progenitor cells isolated from fetal liver cells (FLCs). We revealed that hnRNP K-overexpression alters self-renewal capacity and differentiation potential of these cells in vitro. Such findings were recapitulated in vivo, as murine recipients of hnRNP K-overexpressing FLCs developed fatal myeloproliferative phenotypes.
To elucidate mechanisms by which hnRNP K overexpression causes myeloid neoplasia, we took an unbiased approach utilizing RNA-immunoprecipitation sequencing (fRIP-Seq). Among RNA transcripts interacting with hnRNP K was RUNX1—a pivotal transcriptional regulator of definitive hematopoiesis commonly mutated or translocated in AML. Consensus hnRNP K binding sites were identified in the 5’ UTR and near the 3’ splice site in intron 5-6 of RUNX1. Fluorescence anisotropy studies confirmed these interactions were direct, and abrogated when hnRNP K binding sites within RUNX1 were mutated. Manipulating hnRNP K expression in human cell lines and murine FLCs substantially altered RUNX1 splicing surrounding exon 6. RNA-sequencing of FLCs confirmed these data, exposing RUNX1 as a significantly differentially spliced entity in the context of hnRNP K overexpression. Importantly, the protein product of this spliced product (RUNX1ΔEx6) exhibited disparate transcriptional activity in reporter assays compared to full-length RUNX1. Furthermore, we identified KH3 as the hnRNP K domain most critical for these splicing alterations; deletion of KH3 markedly abrogated hnRNP K overexpression phenotypes in vitro.
In sum, we established hnRNP K as an oncogene in myeloid leukemia that binds RUNX1 RNA, altering its splicing and subsequent transcriptional activity. These findings shed light on a mechanism of myeloid leukemogenesis, paving the way for drug discovery efforts to improve outcomes for patients with this disease.
"]},{"key":"dc:title","label":"Title","values":["Heterogeneous Nuclear Ribonucleoprotein K (Hnrnp K) Overexpression and Its Interaction With Runx1 Rna In Acute Myeloid Leukemia"]}]}],"canonical_facts":{"dc:contributor":["Sean Post, Ph.D.","Scott Evans, M.D.","Marina Konopleva, M.D., Ph.D."],"dc:creator":["Aitken, Marisa","https://orcid.org/0000-0002-7937-531X
"],"dc:date.available":["2021-04-16T07:00:00Z"],"dc:description.abstract":["Acute myeloid leukemia (AML) is an often devastating hematologic malignancy with 5-year overall survival lingering near 20%. Acquiring a deeper understanding of molecular underpinnings of leukemogenesis will provide a basis for developing more effective therapeutic strategies for patients with AML.
Here, we identified overexpression of hnRNP K as a recurrent abnormality in a subset (~20%) of AML patients. High levels of this RNA-binding protein associated with inferior clinical outcomes in de novo AML. Thus, to evaluate its putative oncogenic capacity in myeloid disease, we overexpressed hnRNP K in murine hematopoietic stem and progenitor cells isolated from fetal liver cells (FLCs). We revealed that hnRNP K-overexpression alters self-renewal capacity and differentiation potential of these cells in vitro. Such findings were recapitulated in vivo, as murine recipients of hnRNP K-overexpressing FLCs developed fatal myeloproliferative phenotypes.
To elucidate mechanisms by which hnRNP K overexpression causes myeloid neoplasia, we took an unbiased approach utilizing RNA-immunoprecipitation sequencing (fRIP-Seq). Among RNA transcripts interacting with hnRNP K was RUNX1—a pivotal transcriptional regulator of definitive hematopoiesis commonly mutated or translocated in AML. Consensus hnRNP K binding sites were identified in the 5’ UTR and near the 3’ splice site in intron 5-6 of RUNX1. Fluorescence anisotropy studies confirmed these interactions were direct, and abrogated when hnRNP K binding sites within RUNX1 were mutated. Manipulating hnRNP K expression in human cell lines and murine FLCs substantially altered RUNX1 splicing surrounding exon 6. RNA-sequencing of FLCs confirmed these data, exposing RUNX1 as a significantly differentially spliced entity in the context of hnRNP K overexpression. Importantly, the protein product of this spliced product (RUNX1ΔEx6) exhibited disparate transcriptional activity in reporter assays compared to full-length RUNX1. Furthermore, we identified KH3 as the hnRNP K domain most critical for these splicing alterations; deletion of KH3 markedly abrogated hnRNP K overexpression phenotypes in vitro.
In sum, we established hnRNP K as an oncogene in myeloid leukemia that binds RUNX1 RNA, altering its splicing and subsequent transcriptional activity. These findings shed light on a mechanism of myeloid leukemogenesis, paving the way for drug discovery efforts to improve outcomes for patients with this disease.
"],"dc:identifier":["https://digitalcommons.library.tmc.edu/utgsbs_dissertations/994"],"dc:subject":["hnRNP K","RNA binding proteins","RNA splicing","AML","myeloproliferative disease","mouse model","Animal Experimentation and Research","Cancer Biology","Cell Biology","Genetics","Hemic and Lymphatic Diseases","Immune System Diseases","Immunopathology","Medicine and Health Sciences"],"dc:title":["Heterogeneous Nuclear Ribonucleoprotein K (Hnrnp K) Overexpression and Its Interaction With Runx1 Rna In Acute Myeloid Leukemia"],"thesis:degree_level":["Dissertation (PhD)"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T05:50:31Z"}