DOCSLIB.ORG

Supraphysiologic Levels of the AML1-ETO Isoform Ae9a Are Essential for Transformation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Supraphysiologic Levels of the AML1-ETO Isoform Ae9a Are Essential for Transformation Supraphysiologic levels of the AML1-ETO isoform AE9a are essential for transformation Kevin A. Linka, Shan Lina, Mahesh Shresthaa, Melissa Bowmana, Mark Wunderlicha, Clara D. Bloomfieldb, Gang Huanga,c, and James C. Mulloya,1 aDivision of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229; bThe Ohio State University Comprehensive Cancer Center, Columbus, OH 43210; and cDivision of Pathology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229 Edited by Dennis A. Carson, University of California, San Diego, La Jolla, CA, and approved June 17, 2016 (received for review December 8, 2015) Chromosomal translocation 8;21 is found in 40% of the FAB M2 sub- of t(8;21) disease (25–27). We have also established human-based type of acute myeloid leukemia (AML). The resultant in-frame fusion models using the CBFB-MYH11 and MLL-AF9 fusion proteins (28– protein AML1-ETO (AE) acts as an initiating oncogene for leukemia 30). These models seem to be faithful representations of human dis- development. AE immortalizes human CD34+ cord blood cells in long- ease at multiple levels, demonstrating a high degree of relevance to term culture. We assessed the transforming properties of the alterna- primary patient samples in a tractable model system. In the present tively spliced AE isoform AE9a (or alternative splicing at exon 9), study, we examined the functional consequences of AE9a expression in which is fully transforming in a murine retroviral model, in human comparison with full-length AE in human HSPCs. Our data identify cord blood cells. Full activity was realized only upon increased fusion oncogene expression level as a critical parameter for AE9a function. protein expression. This effect was recapitulated in the AE9a murine Results AML model. Cotransduction of AE and AE9a resulted in a strong se- + lective pressure for AE-expressing cells. In the context of AE, AE9a did AE9a Promotes Self-Renewal of Human CD34 Hematopoietic Cells. not show selection for increased expression, affirming observations of We examined the function of the AML1-ETO isoform AE9a (and the Δ + human t(8;21) patient samples where full-length AE is the dominant AE9a 2 mutant as a negative control) in human CD34 umbilical cord protein detected. Mechanistically, AE9a showed defective transcrip- blood (UCB) cells (Fig. 1A). Cells were transduced with MSCV-HA-AE- IRES-Thy1.1-,AE9a-,orAE9aΔ2-expressing retrovirus and sorted for tional regulation of AE target genes that was partially corrected at Thy1.1 expression. AE9a partially recapitulated the effects of AE tran- high expression. Together, these results bring an additional perspec- scriptional activation and repression although most target genes were tive to our understanding of AE function and highlight the contribu- regulatedtoalesserextentcomparedwithAE,includingCEBPA,SPI1, tion of oncogene expression level in t(8;21) experimental models. OGG1, CDKN1A, and MPL, indicating some loss of function for AE9a + (Fig. 1B). AE9a extended cell growth and maintained a CD34 pop- oncogene dosage | AML | isoform | transformation ulation throughout the lifespan of the cultures, similar to the effects of AE (Fig. 1 C and D). AE9a cultures consistently trended toward a higher he t(8;21)(q22;q22) chromosomal translocation comprises the percentage of CD34-expressing cells over time, unlike AE cultures, TN-terminal DNA binding domain of AML1 (RUNX1) and which showed relatively constant levels of CD34-expressing cells. How- nearly the entire ETO (RUNX1T1) gene, forming the fusion ever, this selection was not advantageous, given the similar growth ki- protein AML1-ETO (AE) (1, 2). Conditional expression and netics of the cells. As expected, deletion of NHR2 in the context of AE9a transduction–transplantation approaches have demonstrated that resulted in a fully nonfunctional mutant, in line with previous findings expression of AE provides self-renewal signaling to hematopoietic (31). AE and AE9a dramatically repressed erythroid colony formation stem/progenitor cells (HSPCs) but does not induce transformation and showed a myeloid colony bias, with significantly fewer colonies in the absence of additional cooperating events (3–7). Several such formed overall compared with controls (Fig. 1E). CD34-expressing cells cooperating events have been identified, including overexpression were maintained in colony-forming unit (cfu) assays, consistent with colony-replating potential for both AE and AE9a (Fig. 1 E and F). of WT1, mutant c-KIT, TEL-PDGFRB, FLT3-ITD, loss of p21, – The ability of the AE9a mutant to transform murine HSPC to AML and treatment with the DNA-damaging agent ENU (6, 8 13). prompted examination of the engraftment and transformation potential Conversely, C-terminal truncation of AE through frameshift muta- of human cells expressing AE9a in comparison with full-length AE. tion (AML1-ETOtr) or alternative splicing at exon 9 (AE9a) leads Cells were transplanted by intrafemoral injection into immunodeficient to acute myeloid leukemia (AML) transformation of murine HSPCs −/− + NOD/SCID IL2RG (NSG) mice transgenic for the human cytokine (14, 15). The AE9a transcript is expressed in ∼70% of t(8;21) AML patients, along with full-length AE (14, 16). AE9a lacks the con- Significance served ETO domains NHR3/4, whose interaction with corepressor proteins such as N-CoR/SMRT and HDACs is important for tran- The AE9a protein (alternative splicing at exon 9) is often used to scriptional repression (17–20). Despite maintaining some corepres- model t(8;21) leukemia. Our study demonstrates that increased sor interaction, AE9a has greatly diminished N-CoR and SMRT oncogene dosage is a critical parameter of AE9a transformation, interaction and is a much less potent transcriptional repressor likely as a result of impaired transcriptional regulation of AML1-ETO than full-length AE (17, 19–22). The AML1-ETOtr mutant showed target genes. This insight could assist in identifying those down- altered regulation of cell cycle proteins, thereby providing a stream genes most critical for t(8;21)-associated transformation. proliferative advantage in K562 cells relative to AE (15). Re- cently, DeKelver et al. showed that the interaction between N-CoR Author contributions: K.A.L., S.L., and J.C.M. designed research; K.A.L., S.L., M.S., M.B., and and the NHR4 domain plays an inhibitory role in leukemia de- M.W. performed research; C.D.B. and G.H. contributed new reagents/analytic tools; K.A.L., MEDICAL SCIENCES velopment in the context of full-length AE (23). However, analysis S.L., M.S., M.W., and J.C.M. analyzed data; and K.A.L., S.L., and J.C.M. wrote the paper. of a small cohort of t(8;21) patient samples did not detect any The authors declare no conflict of interest. mutations in the NHR4 domain, indicating that such mutations This article is a PNAS Direct Submission. are probably very infrequent (24). The role of AE9a in the genesis Data deposition: The data reported in this paper have been deposited in the Gene Ex- of t(8;21)-associated AML is currently unclear. pression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. GSE84513). + Previously, we showed that expression of AE in human CD34 1To whom correspondence should be addressed. Email: [email protected]. HSPCs promotes enhanced self-renewal and long-term expansion of This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. + CD34 progenitors, making this model a valuable preleukemia model 1073/pnas.1524225113/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1524225113 PNAS | August 9, 2016 | vol. 113 | no. 32 | 9075–9080 Downloaded by guest on October 2, 2021 AB protein synthesis inhibitor cycloheximide (Fig. S1A). Protein turnover for AE9a was actually more rapid than for full-length AE, suggesting that increased protein stability was not contributing to high AE9a ex- pression. Given the decreased ability of AE9a to efficiently repress AE target genes, it was possible that AE9a does not repress expression from the murine stem cell virus (MSCV) promoter [which contains a core binding factor (CBF) recognition site] whereas AE does. However, AE C D and AE9a showed similar activity on an MSCV luciferase reporter, suggesting that hyperactivation of the MSCV promoter by AE9a is unlikely to be driving the increased AE9a expression (Fig. S1B). High AE9a Expression Is Essential for Establishment of Long-Term Human E Cultures. To determine whether high expression is necessary for AE9a function, human UCB cells transduced with vector, AE, or AE9a were sorted for expression of the vector-encoded Thy1.1 surface marker immediately after transduction. Fractions with the lowest (30% posi- tive) and highest (30% positive) expression were sorted, and cell growth was monitored on a weekly basis (Fig. 3 A and B). AE-low cells showed robust growth, and expression analysis at week 5 showed that F G AE-low cultures reached a median level of AE expression, based on both AE protein and Thy1.1 expression (Fig. 3A and Fig. S2). AE-high cells initially showed a lag in proliferation (Fig. 3B, Inset), possibly due to an up-regulation of the p21 cell cycle inhibitor (Fig. 3A), but re- covered growth potential after 2–3 wk, presumably due to outgrowth of clones expressing lower levels of AE (Fig. 3 A and B). In contrast, AE9a-high cells had excellent proliferative ability but, interestingly, Fig. 1. AE9a extends CD34+ UCB culture life, retains CD34 progenitor marker maintained or even increased AE9a levels over time (Fig. 3 A and B and expression, and engrafts immunodeficient mice. (A) Schematic of HA-tagged Fig. S2). Strikingly, AE9a-low cells showed no enhanced proliferation + AE, AE9a, or AE9aΔ2. CD34 umbilical cord blood (UCB) cells were transduced with MIT-HA-AE, -AE9a, or -AE9aΔ2, or empty vector and sorted for Thy1.1 surface marker expression. (B–G) Quantitative PCR (QPCR) analysis was per- formed on week 1 transduced AE and mutant cultures.
Load more

Recommended publications
  • Variations in Microrna-25 Expression Influence the Severity of Diabetic
    BASIC RESEARCH www.jasn.org Variations in MicroRNA-25 Expression Influence the Severity of Diabetic Kidney Disease † † † Yunshuang Liu,* Hongzhi Li,* Jieting Liu,* Pengfei Han, Xuefeng Li, He Bai,* Chunlei Zhang,* Xuelian Sun,* Yanjie Teng,* Yufei Zhang,* Xiaohuan Yuan,* Yanhui Chu,* and Binghai Zhao* *Heilongjiang Key Laboratory of Anti-Fibrosis Biotherapy, Medical Research Center, Heilongjiang, People’s Republic of China; and †Clinical Laboratory of Hong Qi Hospital, Mudanjiang Medical University, Heilongjiang, People’s Republic of China ABSTRACT Diabetic nephropathy is characterized by persistent albuminuria, progressive decline in GFR, and second- ary hypertension. MicroRNAs are dysregulated in diabetic nephropathy, but identification of the specific microRNAs involved remains incomplete. Here, we show that the peripheral blood from patients with diabetes and the kidneys of animals with type 1 or 2 diabetes have low levels of microRNA-25 (miR-25) compared with those of their nondiabetic counterparts. Furthermore, treatment with high glucose decreased the expression of miR-25 in cultured kidney cells. In db/db mice, systemic administration of an miR-25 agomir repressed glomerular fibrosis and reduced high BP. Notably, knockdown of miR-25 in normal mice by systemic administration of an miR-25 antagomir resulted in increased proteinuria, extracellular matrix accumulation, podocyte foot process effacement, and hypertension with renin-angiotensin system activation. However, excessive miR-25 did not cause kidney dysfunction in wild-type mice. RNA sequencing showed the alteration of miR-25 target genes in antagomir-treated mice, including the Ras-related gene CDC42. In vitro,cotrans- fection with the miR-25 antagomir repressed luciferase activity from a reporter construct containing the CDC42 39 untranslated region.
    [Show full text]
  • (P -Value<0.05, Fold Change≥1.4), 4 Vs. 0 Gy Irradiation
    Table S1: Significant differentially expressed genes (P -Value<0.05, Fold Change≥1.4), 4 vs. 0 Gy irradiation Genbank Fold Change P -Value Gene Symbol Description Accession Q9F8M7_CARHY (Q9F8M7) DTDP-glucose 4,6-dehydratase (Fragment), partial (9%) 6.70 0.017399678 THC2699065 [THC2719287] 5.53 0.003379195 BC013657 BC013657 Homo sapiens cDNA clone IMAGE:4152983, partial cds. [BC013657] 5.10 0.024641735 THC2750781 Ciliary dynein heavy chain 5 (Axonemal beta dynein heavy chain 5) (HL1). 4.07 0.04353262 DNAH5 [Source:Uniprot/SWISSPROT;Acc:Q8TE73] [ENST00000382416] 3.81 0.002855909 NM_145263 SPATA18 Homo sapiens spermatogenesis associated 18 homolog (rat) (SPATA18), mRNA [NM_145263] AA418814 zw01a02.s1 Soares_NhHMPu_S1 Homo sapiens cDNA clone IMAGE:767978 3', 3.69 0.03203913 AA418814 AA418814 mRNA sequence [AA418814] AL356953 leucine-rich repeat-containing G protein-coupled receptor 6 {Homo sapiens} (exp=0; 3.63 0.0277936 THC2705989 wgp=1; cg=0), partial (4%) [THC2752981] AA484677 ne64a07.s1 NCI_CGAP_Alv1 Homo sapiens cDNA clone IMAGE:909012, mRNA 3.63 0.027098073 AA484677 AA484677 sequence [AA484677] oe06h09.s1 NCI_CGAP_Ov2 Homo sapiens cDNA clone IMAGE:1385153, mRNA sequence 3.48 0.04468495 AA837799 AA837799 [AA837799] Homo sapiens hypothetical protein LOC340109, mRNA (cDNA clone IMAGE:5578073), partial 3.27 0.031178378 BC039509 LOC643401 cds. [BC039509] Homo sapiens Fas (TNF receptor superfamily, member 6) (FAS), transcript variant 1, mRNA 3.24 0.022156298 NM_000043 FAS [NM_000043] 3.20 0.021043295 A_32_P125056 BF803942 CM2-CI0135-021100-477-g08 CI0135 Homo sapiens cDNA, mRNA sequence 3.04 0.043389246 BF803942 BF803942 [BF803942] 3.03 0.002430239 NM_015920 RPS27L Homo sapiens ribosomal protein S27-like (RPS27L), mRNA [NM_015920] Homo sapiens tumor necrosis factor receptor superfamily, member 10c, decoy without an 2.98 0.021202829 NM_003841 TNFRSF10C intracellular domain (TNFRSF10C), mRNA [NM_003841] 2.97 0.03243901 AB002384 C6orf32 Homo sapiens mRNA for KIAA0386 gene, partial cds.
    [Show full text]
  • Genetic Testing for Acute Myeloid Leukemia AHS-M2062
    Corporate Medical Policy Genetic Testing for Acute Myeloid Leukemia AHS-M2062 File Name: genetic_testing_for_acute_myeloid_leukemia Origination: 1/1/2019 Last CAP Review: 8/2021 Next CAP Review: 8/2022 Last Review: 8/2021 Description of Procedure or Service Acute myeloid leukemia (AML) is characterized by large numbers of abnormal, immature myeloid cells in the bone marrow and peripheral blood resulting from genetic changes in hematopoietic precursor cells which disrupt normal hematopoietic growth and differentiation (Stock, 2020). Related Policies: Genetic Cancer Susceptibility Using Next Generation Sequencing AHS-M2066 Molecular Panel Testing of Cancers to Identify Targeted Therapy AHS-M2109 Serum Tumor Markers for Malignancies AHS-G2124 Minimal Residual Disease (MRD) AHS- M2175 ***Note: This Medical Policy is complex and technical. For questions concerning the technical language and/or specific clinical indications for its use, please consult your physician. Policy BCBSNC will provide coverage for genetic testing for acute myeloid leukemia when it is determined to be medically necessary because the medical criteria and guidelines shown below are met. Benefits Application This medical policy relates only to the services or supplies described herein. Please refer to the Member's Benefit Booklet for availability of benefits. Member's benefits may vary according to benefit design; therefore member benefit language should be reviewed before applying the terms of this medical policy. When Genetic Testing for Acute Myeloid Leukemia is covered The use of genetic testing for acute myeloid leukemia is considered medically necessary for the following: A. Genetic testing for FLT3 internal tandem duplication and tyrosine kinase domain mutations (ITD and TKD), IDH1, IDH2, TET2, WT1, DNMT3A, ASXL1 and/or TP53 in adult and pediatric patients with suspected or confirmed AML of any type for prognostic and/or therapeutic purposes.
    [Show full text]
  • Application of Microrna Database Mining in Biomarker Discovery and Identification of Therapeutic Targets for Complex Disease
    Article Application of microRNA Database Mining in Biomarker Discovery and Identification of Therapeutic Targets for Complex Disease Jennifer L. Major, Rushita A. Bagchi * and Julie Pires da Silva * Department of Medicine, Division of Cardiology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA; [email protected] * Correspondence: [email protected] (R.A.B.); [email protected] (J.P.d.S.) Supplementary Tables Methods Protoc. 2021, 4, 5. https://doi.org/10.3390/mps4010005 www.mdpi.com/journal/mps Methods Protoc. 2021, 4, 5. https://doi.org/10.3390/mps4010005 2 of 25 Table 1. List of all hsa-miRs identified by Human microRNA Disease Database (HMDD; v3.2) analysis. hsa-miRs were identified using the term “genetics” and “circulating” as input in HMDD. Targets CAD hsa-miR-1 Targets IR injury hsa-miR-423 Targets Obesity hsa-miR-499 hsa-miR-146a Circulating Obesity Genetics CAD hsa-miR-423 hsa-miR-146a Circulating CAD hsa-miR-149 hsa-miR-499 Circulating IR Injury hsa-miR-146a Circulating Obesity hsa-miR-122 Genetics Stroke Circulating CAD hsa-miR-122 Circulating Stroke hsa-miR-122 Genetics Obesity Circulating Stroke hsa-miR-26b hsa-miR-17 hsa-miR-223 Targets CAD hsa-miR-340 hsa-miR-34a hsa-miR-92a hsa-miR-126 Circulating Obesity Targets IR injury hsa-miR-21 hsa-miR-423 hsa-miR-126 hsa-miR-143 Targets Obesity hsa-miR-21 hsa-miR-223 hsa-miR-34a hsa-miR-17 Targets CAD hsa-miR-223 hsa-miR-92a hsa-miR-126 Targets IR injury hsa-miR-155 hsa-miR-21 Circulating CAD hsa-miR-126 hsa-miR-145 hsa-miR-21 Targets Obesity hsa-mir-223 hsa-mir-499 hsa-mir-574 Targets IR injury hsa-mir-21 Circulating IR injury Targets Obesity hsa-mir-21 Targets CAD hsa-mir-22 hsa-mir-133a Targets IR injury hsa-mir-155 hsa-mir-21 Circulating Stroke hsa-mir-145 hsa-mir-146b Targets Obesity hsa-mir-21 hsa-mir-29b Methods Protoc.
    [Show full text]
  • Supplementary Materials and Tables a and B
    SUPPLEMENTARY MATERIAL 1 Table A. Main characteristics of the subset of 23 AML patients studied by high-density arrays (subset A) WBC BM blasts MYST3- MLL Age/Gender WHO / FAB subtype Karyotype FLT3-ITD NPM status (x109/L) (%) CREBBP status 1 51 / F M4 NA 21 78 + - G A 2 28 / M M4 t(8;16)(p11;p13) 8 92 + - G G 3 53 / F M4 t(8;16)(p11;p13) 27 96 + NA G NA 4 24 / M PML-RARα / M3 t(15;17) 5 90 - - G G 5 52 / M PML-RARα / M3 t(15;17) 1.5 75 - - G G 6 31 / F PML-RARα / M3 t(15;17) 3.2 89 - - G G 7 23 / M RUNX1-RUNX1T1 / M2 t(8;21) 38 34 - + ND G 8 52 / M RUNX1-RUNX1T1 / M2 t(8;21) 8 68 - - ND G 9 40 / M RUNX1-RUNX1T1 / M2 t(8;21) 5.1 54 - - ND G 10 63 / M CBFβ-MYH11 / M4 inv(16) 297 80 - - ND G 11 63 / M CBFβ-MYH11 / M4 inv(16) 7 74 - - ND G 12 59 / M CBFβ-MYH11 / M0 t(16;16) 108 94 - - ND G 13 41 / F MLLT3-MLL / M5 t(9;11) 51 90 - + G R 14 38 / F M5 46, XX 36 79 - + G G 15 76 / M M4 46 XY, der(10) 21 90 - - G NA 16 59 / M M4 NA 29 59 - - M G 17 26 / M M5 46, XY 295 92 - + G G 18 62 / F M5 NA 67 88 - + M A 19 47 / F M5 del(11q23) 17 78 - + M G 20 50 / F M5 46, XX 61 59 - + M G 21 28 / F M5 46, XX 132 90 - + G G 22 30 / F AML-MD / M5 46, XX 6 79 - + M G 23 64 / M AML-MD / M1 46, XY 17 83 - + M G WBC: white blood cell.
    [Show full text]
  • Maintenance of Mammary Epithelial Phenotype by Transcription Factor Runx1 Through Mitotic Gene Bookmarking Joshua Rose University of Vermont
    University of Vermont ScholarWorks @ UVM Graduate College Dissertations and Theses Dissertations and Theses 2019 Maintenance Of Mammary Epithelial Phenotype By Transcription Factor Runx1 Through Mitotic Gene Bookmarking Joshua Rose University of Vermont Follow this and additional works at: https://scholarworks.uvm.edu/graddis Part of the Biochemistry Commons, and the Genetics and Genomics Commons Recommended Citation Rose, Joshua, "Maintenance Of Mammary Epithelial Phenotype By Transcription Factor Runx1 Through Mitotic Gene Bookmarking" (2019). Graduate College Dissertations and Theses. 998. https://scholarworks.uvm.edu/graddis/998 This Thesis is brought to you for free and open access by the Dissertations and Theses at ScholarWorks @ UVM. It has been accepted for inclusion in Graduate College Dissertations and Theses by an authorized administrator of ScholarWorks @ UVM. For more information, please contact [email protected]. MAINTENANCE OF MAMMARY EPITHELIAL PHENOTYPE BY TRANSCRIPTION FACTOR RUNX1 THROUGH MITOTIC GENE BOOKMARKING A Thesis Presented by Joshua Rose to The Faculty of the Graduate College of The University of Vermont In Partial Fulfillment of the Requirements for the Degree of Master of Science Specializing in Cellular, Molecular, and Biomedical Sciences January, 2019 Defense Date: November 12, 2018 Thesis Examination Committee: Sayyed Kaleem Zaidi, Ph.D., Advisor Gary Stein, Ph.D., Advisor Seth Frietze, Ph.D., Chairperson Janet Stein, Ph.D. Jonathan Gordon, Ph.D. Cynthia J. Forehand, Ph.D. Dean of the Graduate College ABSTRACT Breast cancer arises from a series of acquired mutations that disrupt normal mammary epithelial homeostasis and create multi-potent cancer stem cells that can differentiate into clinically distinct breast cancer subtypes. Despite improved therapies and advances in early detection, breast cancer remains the leading diagnosed cancer in women.
    [Show full text]
  • Castration Delays Epigenetic Aging and Feminizes DNA
    RESEARCH ARTICLE Castration delays epigenetic aging and feminizes DNA methylation at androgen- regulated loci Victoria J Sugrue1, Joseph Alan Zoller2, Pritika Narayan3, Ake T Lu4, Oscar J Ortega-Recalde1, Matthew J Grant3, C Simon Bawden5, Skye R Rudiger5, Amin Haghani4, Donna M Bond1, Reuben R Hore6, Michael Garratt1, Karen E Sears7, Nan Wang8, Xiangdong William Yang8,9, Russell G Snell3, Timothy A Hore1†*, Steve Horvath4†* 1Department of Anatomy, University of Otago, Dunedin, New Zealand; 2Department of Biostatistics, Fielding School of Public Health, University of California, Los Angeles, Los Angeles, United States; 3Applied Translational Genetics Group, School of Biological Sciences, Centre for Brain Research, The University of Auckland, Auckland, New Zealand; 4Department of Human Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, United States; 5Livestock and Farming Systems, South Australian Research and Development Institute, Roseworthy, Australia; 6Blackstone Hill Station, Becks, RD2, Omakau, New Zealand; 7Department of Ecology and Evolutionary Biology, UCLA, Los Angeles, United States; 8Center for Neurobehavioral Genetics, Semel Institute for Neuroscience and Human Behavior, University of California, Los Angeles (UCLA), Los Angeles, United States; 9Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine at UCLA, Los Angeles, United States *For correspondence: Abstract In mammals, females generally live longer than males. Nevertheless, the mechanisms [email protected] (TAH); underpinning sex-dependent longevity are currently unclear. Epigenetic clocks are powerful [email protected] (SH) biological biomarkers capable of precisely estimating chronological age and identifying novel †These authors contributed factors influencing the aging rate using only DNA methylation data. In this study, we developed the equally to this work first epigenetic clock for domesticated sheep (Ovis aries), which can predict chronological age with a median absolute error of 5.1 months.
    [Show full text]
  • Engineered Type 1 Regulatory T Cells Designed for Clinical Use Kill Primary
    ARTICLE Acute Myeloid Leukemia Engineered type 1 regulatory T cells designed Ferrata Storti Foundation for clinical use kill primary pediatric acute myeloid leukemia cells Brandon Cieniewicz,1* Molly Javier Uyeda,1,2* Ping (Pauline) Chen,1 Ece Canan Sayitoglu,1 Jeffrey Mao-Hwa Liu,1 Grazia Andolfi,3 Katharine Greenthal,1 Alice Bertaina,1,4 Silvia Gregori,3 Rosa Bacchetta,1,4 Norman James Lacayo,1 Alma-Martina Cepika1,4# and Maria Grazia Roncarolo1,2,4# Haematologica 2021 Volume 106(10):2588-2597 1Department of Pediatrics, Division of Stem Cell Transplantation and Regenerative Medicine, Stanford School of Medicine, Stanford, CA, USA; 2Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford School of Medicine, Stanford, CA, USA; 3San Raffaele Telethon Institute for Gene Therapy, Milan, Italy and 4Center for Definitive and Curative Medicine, Stanford School of Medicine, Stanford, CA, USA *BC and MJU contributed equally as co-first authors #AMC and MGR contributed equally as co-senior authors ABSTRACT ype 1 regulatory (Tr1) T cells induced by enforced expression of interleukin-10 (LV-10) are being developed as a novel treatment for Tchemotherapy-resistant myeloid leukemias. In vivo, LV-10 cells do not cause graft-versus-host disease while mediating graft-versus-leukemia effect against adult acute myeloid leukemia (AML). Since pediatric AML (pAML) and adult AML are different on a genetic and epigenetic level, we investigate herein whether LV-10 cells also efficiently kill pAML cells. We show that the majority of primary pAML are killed by LV-10 cells, with different levels of sensitivity to killing. Transcriptionally, pAML sensitive to LV-10 killing expressed a myeloid maturation signature.
    [Show full text]
  • The Genetic Landscape and Clonal Evolution of Breast Cancer Resistance to Palbociclib Plus Fulvestrant in the PALOMA-3 Trial
    Published OnlineFirst September 11, 2018; DOI: 10.1158/2159-8290.CD-18-0264 RESEARCH ARTICLE The Genetic Landscape and Clonal Evolution of Breast Cancer Resistance to Palbociclib plus Fulvestrant in the PALOMA-3 Trial Ben O’Leary1,2, Rosalind J. Cutts1, Yuan Liu3, Sarah Hrebien1, Xin Huang3, Kerry Fenwick4, Fabrice André5, Sibylle Loibl6, Sherene Loi7, Isaac Garcia-Murillas1, Massimo Cristofanilli8, Cynthia Huang Bartlett3, and Nicholas C. Turner1,2 Downloaded from cancerdiscovery.aacrjournals.org on September 24, 2021. © 2018 American Association for Cancer Research. Published OnlineFirst September 11, 2018; DOI: 10.1158/2159-8290.CD-18-0264 ABSTRACT CDK4/6 inhibition with endocrine therapy is now a standard of care for advanced estrogen receptor–positive breast cancer. Mechanisms of CDK4/6 inhibitor resist- ance have been described preclinically, with limited evidence from clinical samples. We conducted paired baseline and end-of-treatment circulating tumor DNA sequencing from 195 patients in the PALOMA-3 randomized phase III trial of palbociclib plus fulvestrant versus placebo plus fulvestrant. We show that clonal evolution occurs frequently during treatment, refl ecting substantial subclonal complexity in breast cancer that has progressed after prior endocrine therapy. RB1 mutations emerged only in the palbociclib plus fulvestrant arm and in a minority of patients (6/127, 4.7%, P = 0.041). New driver mutations emerged in PIK3CA (P = 0.00069) and ESR1 after treatment in both arms, in particular ESR1 Y537S (P = 0.0037). Evolution of driver gene mutations was uncommon in patients progressing early on palbociclib plus fulvestrant but common in patients progressing later on treatment. These fi nd- ings inform future treatment strategies to address resistance to palbociclib plus fulvestrant.
    [Show full text]
  • Interplay Between Cofactors and Transcription Factors in Hematopoiesis and Hematological Malignancies
    Signal Transduction and Targeted Therapy www.nature.com/sigtrans REVIEW ARTICLE OPEN Interplay between cofactors and transcription factors in hematopoiesis and hematological malignancies Zi Wang 1,2, Pan Wang2, Yanan Li2, Hongling Peng1, Yu Zhu2, Narla Mohandas3 and Jing Liu2 Hematopoiesis requires finely tuned regulation of gene expression at each stage of development. The regulation of gene transcription involves not only individual transcription factors (TFs) but also transcription complexes (TCs) composed of transcription factor(s) and multisubunit cofactors. In their normal compositions, TCs orchestrate lineage-specific patterns of gene expression and ensure the production of the correct proportions of individual cell lineages during hematopoiesis. The integration of posttranslational and conformational modifications in the chromatin landscape, nucleosomes, histones and interacting components via the cofactor–TF interplay is critical to optimal TF activity. Mutations or translocations of cofactor genes are expected to alter cofactor–TF interactions, which may be causative for the pathogenesis of various hematologic disorders. Blocking TF oncogenic activity in hematologic disorders through targeting cofactors in aberrant complexes has been an exciting therapeutic strategy. In this review, we summarize the current knowledge regarding the models and functions of cofactor–TF interplay in physiological hematopoiesis and highlight their implications in the etiology of hematological malignancies. This review presents a deep insight into the physiological and pathological implications of transcription machinery in the blood system. Signal Transduction and Targeted Therapy (2021) ;6:24 https://doi.org/10.1038/s41392-020-00422-1 1234567890();,: INTRODUCTION by their ATPase subunits into four major families, including the Hematopoiesisisacomplexhierarchicaldifferentiationprocessthat SWI/SNF, ISWI, Mi-2/NuRD, and INO80/SWR1 families.
    [Show full text]
  • (NGS) for Primary Endocrine Resistance in Breast Cancer Patients
    Int J Clin Exp Pathol 2018;11(11):5450-5458 www.ijcep.com /ISSN:1936-2625/IJCEP0084102 Original Article Impact of next-generation sequencing (NGS) for primary endocrine resistance in breast cancer patients Ruoyang Li1*, Tiantian Tang1*, Tianli Hui1, Zhenchuan Song1, Fugen Li2, Jingyu Li2, Jiajia Xu2 1Breast Center, Fourth Hospital of Hebei Medical University, Shijiazhuang, China; 2Institute of Precision Medicine, 3D Medicines Inc., Shanghai, China. *Equal contributors. Received August 15, 2018; Accepted September 22, 2018; Epub November 1, 2018; Published November 15, 2018 Abstract: Multiple mechanisms have been detected to account for the acquired resistance to endocrine therapies in breast cancer. In this study we retrospectively studied the mechanism of primary endocrine resistance in estrogen receptor positive (ER+) breast cancer patients by next-generation sequencing (NGS). Tumor specimens and matched blood samples were obtained from 24 ER+ breast cancer patients. Fifteen of them displayed endocrine resistance, including recurrence and/or metastases within 24 months from the beginning of endocrine therapy, and 9 pa- tients remained sensitive to endocrine therapy for more than 5 years. Genomic DNA of tumor tissue was extracted from formalin-fixed paraffin-embedded (FFPE) tumor tissue blocks. Genomic DNA of normal tissue was extracted from peripheral blood mononuclear cells (PBMC). Sequencing libraries for each sample were prepared, followed by target capturing for 372 genes that are frequently rearranged in cancers. Massive parallel
    [Show full text]
  • Investigating Caveolin-Dependent Glucocorticoid Action in Inflammation
    Investigating caveolin-dependent glucocorticoid action in inflammation A thesis submitted to the University of Manchester for the degree of M.Phil in the Faculty of Medicine and Human Sciences 2013 Anna Louise Mary Mannion-Jones School of Medicine 1 Contents I. TABLES AND FIGURES .................................................................................................. 7 II. Abstract .................................................................................................................... 10 III. Declaration ............................................................................................................ 10 IV. Notice of Copyright................................................................................................ 11 1 Introduction .............................................................................................................. 18 1.1 Inflammation ..................................................................................................... 18 1.1.1 The cellular inflammatory response ........................................................... 18 1.1.2 Innate vs. adaptive immunity ..................................................................... 19 1.1.3 Chronic inflammation ................................................................................. 20 1.1.4 Autoimmune disease.................................................................................. 20 1.1.5 Anti-inflammatory actions of glucocorticoids ............................................. 22 1.1.6 The
    [Show full text]