Functional Interaction of BRCA1 and CREBBP in Murine Hematopoiesis

Holmstrom, Sam R.; Wijayatunge, Ranjula; McCrum, Kelly; Mgbemena, Victoria; Ross, Theodora S.

doi:10.1016/j.isci.2019.08.031

Cited by 4 publications

(4 citation statements)

References 31 publications

(65 reference statements)

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“…Use of unspecific or unreliable antibodies for mouse endogenous BRCA1 protein detection occurred in the field in the past three decades. In this study, we found that antibody 287.17 specifically recognizes mouse BRCA1 protein in WB, IP and IF, consistent with reports from recent publications 76 , 77 . We hope our rigorous characterization of BRCA1 antibodies will be helpful to researchers in the BRCA1 field especially those who study BRCA1 in mouse models.…”

Section: Discussionsupporting

confidence: 93%

BRCA1 Antibodies Matter

Yang

Chiang

et al. 2021

Int. J. Biol. Sci.

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Breast cancer susceptibility gene 1 ( BRCA1 ) encodes a tumor suppressor that is frequently mutated in familial breast and ovarian cancer patients. BRCA1 functions in multiple important cellular processes including DNA damage repair, cell cycle checkpoint activation, protein ubiquitination, chromatin remodeling, transcriptional regulation, as well as R-loop formation and apoptosis. A large number of BRCA1 antibodies have been generated and become commercially available over the past three decades, however, many commercial antibodies are poorly characterized and, when widely used, led to unreliable data. In search of reliable and specific BRCA1 antibodies (Abs), particularly antibodies recognizing mouse BRCA1, we performed a rigorous validation of a number of commercially available anti-BRCA1 antibodies, using proper controls in a panel of validation applications, including Western blot (WB), immunoprecipitation (IP), immunoprecipitation-mass spectrometry (IP-MS), chromatin immunoprecipitation (ChIP) and immunofluorescence (IF). Furthermore, we assessed the specificity of these antibodies to detect mouse BRCA1 protein through the use of testis tissue and mouse embryonic fibroblasts (MEFs) from Brca1 +/+ and Brca1 Δ11/Δ11 mice. We find that Ab1, D-9, 07-434 (for recognizing human BRCA1) and 287.17, 440621, BR-64 (for recognizing mouse BRCA1) are specific with high quality performance in the indicated assays. We share these results here with the goal of helping the community combat the common challenges associated with anti-BRCA1 antibody specificity and reproducibility and, hopefully, better understanding BRCA1 functions at cellular and tissue levels.

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Section: Discussionsupporting

confidence: 93%

BRCA1 Antibodies Matter

Yang

Chiang

et al. 2021

Int. J. Biol. Sci.

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“…Flow samples were analyzed using the FACS Canto RUO analyzer (BD Biosciences). Gating schemes for hematopoietic progenitors were performed as previously described ( 17 , 63 , 64 ) using fluorescence minus 1 control. Complete peripheral blood counts were assessed using the HemaVet HV950 with MULTI-TROL Mouse as an equilibration control (Drew Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…For flow experiments, bone marrow and spleen single cells suspensions were created as described (63). For HSC and progenitor analysis cells were incubated with biotinylated lineage specific antibodies (CD3ε, CD4, CD5, CD8a, CD45R, CD11b, Gr-1, Ter119) and fluorophore-conjugated antibodies against progenitor surface markers (Sca1, c-kit, CD16/32, CD34, CD48, CD150).…”

Section: Hematopoietic Analysismentioning

confidence: 99%

BRCA1 and TP53 codeficiency causes a PARP inhibitor–sensitive erythroproliferative neoplasm

Lopez-Perez¹,

Wijayatunge²,

McCrum³

et al. 2022

JCI Insight

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Mutations in the BRCA1 tumor suppressor gene, such as 5382insC (BRCA1 5382insC ), give carriers an increased risk for breast, ovarian, prostate and pancreatic cancers. We have previously reported that, in mice, Brca1 deficiency in the hematopoietic system leads to pancytopenia and, as a result, early lethality. Here we explore the cellular consequences of Brca1 null and BRCA1 5382insC alleles in combination with Trp53 deficiency in the murine hematopoietic system. We find that Brca1 and Trp53 co-deficiency leads to a highly penetrant erythroproliferative disorder that is characterized by hepatosplenomegaly and expanded megakaryocyte erythroid progenitor (MEP) and immature erythroid blast populations. The expanded erythroid progenitor populations in both bone marrow and spleen have the capacity to transmit the disease into secondary mouse recipients, suggesting Brca1 and Trp53 co-deficiency provides a new murine model of hematopoietic neoplasia. This Brca1/Trp53 model replicates Poly (ADP-ribose) polymerase (PARP) inhibitor olaparib sensitivity seen in existing Brca1/Trp53 breast cancer models and has the benefits of monitoring disease progression and drug responses via peripheral blood analyses without sacrificing experimental animals. In addition, this erythroid neoplasia develops much faster than murine breast cancer, allowing for increased efficiency of future preclinical studies.

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“…Crebbp and Brca1 functionally interact to maintain normal hematopoiesis. Crebbp haploinsufficiency in mice is deleterious for the bone marrow microenvironment, which results in myeloproliferation associated with an increase in splenic HSCs as well as a lethal systemic inflammatory disorder [35,36].…”

Section: The Role Of Crebbp/ep300 In Normal Hematopoiesismentioning

confidence: 99%

The Role of CREBBP/EP300 and Its Therapeutic Implications in Hematological Malignancies

Zhu

Wang

et al. 2023

Cancers

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Disordered histone acetylation has emerged as a key mechanism in promoting hematological malignancies. CREB-binding protein (CREBBP) and E1A-binding protein P300 (EP300) are two key acetyltransferases and transcriptional cofactors that regulate gene expression by regulating the acetylation levels of histone proteins and non-histone proteins. CREBBP/EP300 dysregulation and CREBBP/EP300-containing complexes are critical for the initiation, progression, and chemoresistance of hematological malignancies. CREBBP/EP300 also participate in tumor immune responses by regulating the differentiation and function of multiple immune cells. Currently, CREBBP/EP300 are attractive targets for drug development and are increasingly used as favorable tools in preclinical studies of hematological malignancies. In this review, we summarize the role of CREBBP/EP300 in normal hematopoiesis and highlight the pathogenic mechanisms of CREBBP/EP300 in hematological malignancies. Moreover, the research basis and potential future therapeutic implications of related inhibitors were also discussed from several aspects. This review represents an in-depth insight into the physiological and pathological significance of CREBBP/EP300 in hematology.

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Functional Interaction of BRCA1 and CREBBP in Murine Hematopoiesis

Cited by 4 publications

References 31 publications

BRCA1 Antibodies Matter

BRCA1 Antibodies Matter

BRCA1 and TP53 codeficiency causes a PARP inhibitor–sensitive erythroproliferative neoplasm

The Role of CREBBP/EP300 and Its Therapeutic Implications in Hematological Malignancies

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