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A Cultured Leukemia Stem Cell Model Enables Validation of CDK6 as a Stemness Target Against Acute Myeloid Leukemia

Acute myeloid leukemia (AML) represents a group of aggressive hematological malignancies, the clinical management of which is made challenging due to the persistence of rare and therapy resistant leukemia stem cells (LSCs) which serve as a source of disease relapse and poor outcomes. There are currently a paucity of methods to reliably enrich and study LSCs, hindering the development of therapies that specifically target LSCs. In this study, we deeply characterize the OCI-AML8227 culture model, which maintains a functional stemness hierarchy originating from its highly primitive CD34⁺CD38⁻ cells, to elucidate LSC biology and uncover LSC-specific therapeutic vulnerabilities. We analyzed both bulk and single-cell proteomics, transcriptomics, and epigenomics to generate a LSC protein-protein interaction network, which was then integrated with an LSC-focused small molecule screen using this model. From these findings, CDK6 was discovered as a therapeutic vulnerability specific to LSCs, which was validated in findings from the BEAT-AML cohort and a patient-derived xenograft (PDX) panel of AML samples through palbociclib treatment. Taken together, our studies validate CDK6 as a druggable vulnerability in LSCs, and authenticate OCI-AML8227 cells as a LSC target discovery engine.

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Access to data generated by UHN is made available by completing the data access agreement for review by the data access committee and will be granted to qualified investigators for appropriate use.

DATA ACCESS AGREEMENT These terms and conditions govern access to the managed access datasets (details of which are set out in Appendix I) to which the User Institution has requested access. The User Institution agrees to be bound by these terms and conditions. Definitions Authorised Personnel: The individuals at the User Institution to whom the Data Access Committee (DAC) grants access to the Data. This includes the User, the individuals listed in Appendix II and any other individuals for whom the User Institution subsequently requests access to the Data. Details of the initial Authorised Personnel are set out in Appendix II. Data: The managed access datasets to which the User Institution has requested access. Data Producers: The Principal Investigator and the collaborators listed in Appendix I responsible for the development, organisation, and oversight of these Data. External Collaborator: A collaborator of the User, working for an institution other than the User Institution. Project: The project for which the User Institution has requested access to these Data. A description of the Project is set out in Appendix II. Publications: Includes, without limitation, articles published in print journals, electronic journals, reviews, books, posters and other written and verbal presentations of research. Research Participant: An individual whose data form part of these Data. Research Purposes: Shall mean research that is seeking to advance the understanding of genetics and genomics, including the treatment of disorders, and work on statistical methods that may be applied to such research. User: The principal investigator for the Project. User Institution(s): The Institution that has requested access to the Data. University Health Network: The University Health Network (UHN) is the largest clinical and research hospital in Canada. UHN is affiliated with the University of Toronto and is a member of the Toronto Academic Health Science Network. 1. The User Institution agrees to only use these Data for the purpose of the Project (described in Appendix II) and only for Research Purposes. The User Institution further agrees that it will only use these Data for Research Purposes which are within the limitations (if any) set out in Appendix I. 2. The User Institution agrees to preserve, at all times, the confidentiality of these Data. In particular, it undertakes not to use, or attempt to use these Data to compromise or otherwise infringe the confidentiality of information on Research Participants. Without prejudice to the generality of the foregoing, the User Institution agrees to use at least the measures set out in Appendix I to protect these Data. 3. The User Institution agrees to protect the confidentiality of Research Participants in any research papers or publications that they prepare by taking all reasonable care to limit the possibility of identification. 4. The User Institution agrees not to link or combine these Data to other information or archived data available in a way that could re-identify the Research Participants, even if access to that data has been formally granted to the User Institution or is freely available without restriction. 5. The User Institution agrees only to transfer or disclose these Data, in whole or part, or any material derived from these Data, to the Authorised Personnel. Should the User Institution wish to share these Data with an External Collaborator, the External Collaborator must complete a separate application for access to these Data. 6. The User Institution agrees that the Data Producers, and all other parties involved in the creation, funding or protection of these Data: a) make no warranty or representation, express or implied as to the accuracy, quality or comprehensiveness of these Data; b) exclude to the fullest extent permitted by law all liability for actions, claims, proceedings, demands, losses (including but not limited to loss of profit), costs, awards damages and payments made by the Recipient that may arise (whether directly or indirectly) in any way whatsoever from the Recipient’s use of these Data or from the unavailability of, or break in access to, these Data for whatever reason and; c) bear no responsibility for the further analysis or interpretation of these Data. 7. The User Institution agrees to follow the Fort Lauderdale Guidelines (http://www.wellcome.ac.uk/stellent/groups/corporatesite/@policy_communications/documents/web_document/wtd003207.pdf ) and the Toronto Statement (http://www.nature.com/nature/journal/v461/n7261/full/461168a.html). This includes but is not limited to recognising the contribution of the Data Producers and including a proper acknowledgement in all reports or publications resulting from the use of these Data. 8. The User Institution agrees to follow the Publication Policy in Appendix III. This includes respecting the moratorium period for the Data Producers to publish the first peer-reviewed report describing and analysing these Data. 9. The User Institution agrees not to make intellectual property claims on these Data and not to use intellectual property protection in ways that would prevent or block access to, or use of, any element of these Data, or conclusion drawn directly from these Data. 10. The User Institution can elect to perform further research that would add intellectual and resource capital to these data and decide to obtain intellectual property rights on these downstream discoveries. In this case, the User Institution agrees to implement licensing policies that will not obstruct further research and to follow the U.S. National Institutes of Health Best Practices for the Licensing of Genomic Inventions (2005) (https://www.icgc.org/files/daco/NIH_BestPracticesLicensingGenomicInventions_2005_en.pdf ) in conformity with the Organisation for Economic Co-operation and Development Guidelines for the Licensing of the Genetic Inventions (2006) (http://www.oecd.org/science/biotech/36198812.pdf ). 11. The User Institution agrees to destroy/discard the Data held, once it is no longer used for the Project, unless obliged to retain the data for archival purposes in conformity with audit or legal requirements. 12. The User Institution will notify the DAC within 30 days of any changes or departures of Authorised Personnel. 13. The User Institution will notify the DAC prior to any significant changes to the protocol for the Project. 14. The User Institution will notify the DAC as soon as it becomes aware of a breach of the terms or conditions of this agreement. 15. The DAC may terminate this agreement by written notice to the User Institution. If this agreement terminates for any reason, the User Institution will be required to destroy any Data held, including copies and backup copies. This clause does not prevent the User Institution from retaining these data for archival purpose in conformity with audit or legal requirements. 16. The User Institution accepts that it may be necessary for the Data Producers to alter the terms of this agreement from time to time. In the event that changes are required, the Data Producers or their appointed agent will contact the User Institution to inform it of the changes and the User Institution may elect to accept the changes or terminate the agreement. 17. If requested, the User Institution will allow data security and management documentation to be inspected to verify that it is complying with the terms of this agreement. 18. The User Institution agrees to distribute a copy of these terms to the Authorised Personnel. The User Institution will procure that the Authorised Personnel comply with the terms of this agreement. 19. This agreement (and any dispute, controversy, proceedings or claim of whatever nature arising out of this agreement or its formation) shall be construed, interpreted and governed by the laws of the province of Ontario and PHIPA. Agreed for User Institution Signature: Name: Title: Date: Principal Investigator I confirm that I have read and understood this Agreement. Signature: Name: Title: Date: Agreed for The Data Access Committee Signature: Name: Title: Date: APPENDIX I – DATASET DETAILS APPENDIX II ––PROJECT DETAILS APPENDIX III –– PUBLICATION POLICY

Studies are experimental investigations of a particular phenomenon, e.g., case-control studies on a particular trait or cancer research projects reporting matching cancer normal genomes from patients.

Study ID Study Title Study Type
EGAS50000000786 RNASeq
EGAS50000000787 Epigenetics
EGAS50000000788 Epigenetics
EGAS50000000789 RNASeq
  • DAC number
  • Dataset Released

This table displays only public information pertaining to the files in the dataset. If you wish to access this dataset, please submit a request. If you already have access to these data files, please consult the download documentation.

ID File Type Size Quality Report
Located in
EGAF50000277486 fastq.gz 3.7 GB
EGAF50000277487 fastq.gz 3.7 GB
EGAF50000277488 fastq.gz 3.8 GB
EGAF50000277489 fastq.gz 3.4 GB
EGAF50000277490 fastq.gz 3.0 GB
EGAF50000277491 fastq.gz 2.5 GB
EGAF50000277492 fastq.gz 3.8 GB
EGAF50000277493 fastq.gz 3.9 GB
EGAF50000277494 fastq.gz 3.3 GB
EGAF50000277495 fastq.gz 3.4 GB
EGAF50000277496 fastq.gz 3.1 GB
EGAF50000277497 fastq.gz 3.2 GB
EGAF50000277498 fastq.gz 3.6 GB
EGAF50000277499 fastq.gz 3.5 GB
EGAF50000277500 fastq.gz 2.6 GB
EGAF50000277501 fastq.gz 2.6 GB
EGAF50000277502 fastq.gz 2.7 GB
EGAF50000277503 fastq.gz 3.4 GB
EGAF50000277504 fastq.gz 3.6 GB
EGAF50000277505 fastq.gz 3.8 GB
EGAF50000277506 fastq.gz 3.7 GB
EGAF50000277507 fastq.gz 3.7 GB
EGAF50000277508 fastq.gz 3.5 GB
EGAF50000277509 fastq.gz 3.6 GB
EGAF50000277510 fastq.gz 2.9 GB
EGAF50000277511 fastq.gz 3.0 GB
EGAF50000277512 fastq.gz 3.2 GB
EGAF50000277513 fastq.gz 3.3 GB
EGAF50000277514 fastq.gz 3.5 GB
EGAF50000277515 fastq.gz 3.6 GB
EGAF50000277516 fastq.gz 3.4 GB
EGAF50000277517 fastq.gz 2.3 GB
EGAF50000277518 fastq.gz 3.1 GB
EGAF50000277519 fastq.gz 3.3 GB
EGAF50000277520 fastq.gz 848.9 MB
EGAF50000277521 fastq.gz 2.4 GB
EGAF50000277522 fastq.gz 5.3 GB
EGAF50000277523 fastq.gz 793.7 MB
EGAF50000277524 fastq.gz 276.0 MB
EGAF50000277525 fastq.gz 297.9 MB
EGAF50000277526 fastq.gz 846.2 MB
EGAF50000277527 fastq.gz 1.8 GB
EGAF50000277528 fastq.gz 3.6 GB
EGAF50000277529 fastq.gz 3.7 GB
EGAF50000277530 fastq.gz 302.0 MB
EGAF50000277531 fastq.gz 856.8 MB
EGAF50000277532 fastq.gz 1.9 GB
EGAF50000277533 fastq.gz 281.2 MB
EGAF50000277534 fastq.gz 1.2 GB
EGAF50000277535 fastq.gz 2.3 GB
EGAF50000277536 fastq.gz 2.3 GB
EGAF50000277537 fastq.gz 534.6 MB
EGAF50000277538 fastq.gz 1.1 GB
EGAF50000277539 fastq.gz 2.1 GB
EGAF50000277540 fastq.gz 507.0 MB
EGAF50000277541 fastq.gz 2.1 GB
EGAF50000277542 fastq.gz 2.1 GB
EGAF50000277543 fastq.gz 506.0 MB
EGAF50000277544 fastq.gz 1.1 GB
EGAF50000277545 fastq.gz 2.1 GB
60 Files (154.0 GB)