Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology

Yuichi Ishikawa, Manami Maeda, Mithun Pasham, Francois Aguet, Silvia K. Tacheva-Grigorova, Takeshi Masuda, Hai Yi, Sung Uk Lee, Jian Xu, Julie Teruya-Feldstein, Maria Ericsson, Ann Mullally, John Heuser, Tom Kirchhausen, Takahiro Maeda

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Clathrin-dependent endocytosis is an essential cellular process shared by all cell types. Despite this, precisely how endocytosis is regulated in a cell-type-specific manner and how this key pathway functions physiologically or pathophysiologically remain largely unknown. PICALM, which encodes the clathrin adaptor protein PICALM, was originally identified as a component of the CALM/AF10 leukemia oncogene. Here we show, by employing a series of conditional Picalm knockout mice, that PICALM critically regulates transferrin uptake in erythroid cells by functioning as a cell-type-specific regulator of transferrin receptor endocytosis. While transferrin receptor is essential for the development of all hematopoietic lineages, Picalm was dispensable for myeloid and B-lymphoid development. Furthermore, global Picalm inactivation in adult mice did not cause gross defects in mouse fitness, except for anemia and a coat color change. Freeze-etch electron microscopy of primary erythroblasts and live-cell imaging of murine embryonic fibroblasts revealed that Picalm function is required for efficient clathrin coat maturation. We showed that the PICALM PIP2 binding domain is necessary for transferrin receptor endocytosis in erythroblasts and absolutely essential for erythroid development from mouse hematopoietic stem/progenitor cells in an erythroid culture system. We further showed that Picalm deletion entirely abrogated the disease phenotype in a Jak2V617F knock-in murine model of polycythemia vera. Our findings provide new insights into the regulation of cell-type-specific transferrin receptor endocytosis in vivo. They also suggest a new strategy to block cellular uptake of transferrin-bound iron, with therapeutic potential for disorders characterized by inappropriate red blood cell production, such as polycythemia vera.

Original languageEnglish
Pages (from-to)439-451
Number of pages13
JournalHaematologica
Volume100
Issue number4
DOIs
Publication statusPublished - Jan 1 2015

Fingerprint

Vesicular Transport Adaptor Proteins
Polycythemia Vera
Hematopoiesis
Endocytosis
Transferrin Receptors
Erythroblasts
Clathrin
Transferrin
Hematopoietic Stem Cells
Erythroid Cells
Oncogenes
Knockout Mice
Anemia
Electron Microscopy
Leukemia
Iron
Color
Fibroblasts
Erythrocytes
Phenotype

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Ishikawa, Y., Maeda, M., Pasham, M., Aguet, F., Tacheva-Grigorova, S. K., Masuda, T., ... Maeda, T. (2015). Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology. Haematologica, 100(4), 439-451. https://doi.org/10.3324/haematol.2014.119537

Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology. / Ishikawa, Yuichi; Maeda, Manami; Pasham, Mithun; Aguet, Francois; Tacheva-Grigorova, Silvia K.; Masuda, Takeshi; Yi, Hai; Lee, Sung Uk; Xu, Jian; Teruya-Feldstein, Julie; Ericsson, Maria; Mullally, Ann; Heuser, John; Kirchhausen, Tom; Maeda, Takahiro.

In: Haematologica, Vol. 100, No. 4, 01.01.2015, p. 439-451.

Research output: Contribution to journalArticle

Ishikawa, Y, Maeda, M, Pasham, M, Aguet, F, Tacheva-Grigorova, SK, Masuda, T, Yi, H, Lee, SU, Xu, J, Teruya-Feldstein, J, Ericsson, M, Mullally, A, Heuser, J, Kirchhausen, T & Maeda, T 2015, 'Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology', Haematologica, vol. 100, no. 4, pp. 439-451. https://doi.org/10.3324/haematol.2014.119537
Ishikawa Y, Maeda M, Pasham M, Aguet F, Tacheva-Grigorova SK, Masuda T et al. Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology. Haematologica. 2015 Jan 1;100(4):439-451. https://doi.org/10.3324/haematol.2014.119537
Ishikawa, Yuichi ; Maeda, Manami ; Pasham, Mithun ; Aguet, Francois ; Tacheva-Grigorova, Silvia K. ; Masuda, Takeshi ; Yi, Hai ; Lee, Sung Uk ; Xu, Jian ; Teruya-Feldstein, Julie ; Ericsson, Maria ; Mullally, Ann ; Heuser, John ; Kirchhausen, Tom ; Maeda, Takahiro. / Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology. In: Haematologica. 2015 ; Vol. 100, No. 4. pp. 439-451.
@article{09f70dc70a3743bdae2fe9e9ac09c9ff,
title = "Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology",
abstract = "Clathrin-dependent endocytosis is an essential cellular process shared by all cell types. Despite this, precisely how endocytosis is regulated in a cell-type-specific manner and how this key pathway functions physiologically or pathophysiologically remain largely unknown. PICALM, which encodes the clathrin adaptor protein PICALM, was originally identified as a component of the CALM/AF10 leukemia oncogene. Here we show, by employing a series of conditional Picalm knockout mice, that PICALM critically regulates transferrin uptake in erythroid cells by functioning as a cell-type-specific regulator of transferrin receptor endocytosis. While transferrin receptor is essential for the development of all hematopoietic lineages, Picalm was dispensable for myeloid and B-lymphoid development. Furthermore, global Picalm inactivation in adult mice did not cause gross defects in mouse fitness, except for anemia and a coat color change. Freeze-etch electron microscopy of primary erythroblasts and live-cell imaging of murine embryonic fibroblasts revealed that Picalm function is required for efficient clathrin coat maturation. We showed that the PICALM PIP2 binding domain is necessary for transferrin receptor endocytosis in erythroblasts and absolutely essential for erythroid development from mouse hematopoietic stem/progenitor cells in an erythroid culture system. We further showed that Picalm deletion entirely abrogated the disease phenotype in a Jak2V617F knock-in murine model of polycythemia vera. Our findings provide new insights into the regulation of cell-type-specific transferrin receptor endocytosis in vivo. They also suggest a new strategy to block cellular uptake of transferrin-bound iron, with therapeutic potential for disorders characterized by inappropriate red blood cell production, such as polycythemia vera.",
author = "Yuichi Ishikawa and Manami Maeda and Mithun Pasham and Francois Aguet and Tacheva-Grigorova, {Silvia K.} and Takeshi Masuda and Hai Yi and Lee, {Sung Uk} and Jian Xu and Julie Teruya-Feldstein and Maria Ericsson and Ann Mullally and John Heuser and Tom Kirchhausen and Takahiro Maeda",
year = "2015",
month = "1",
day = "1",
doi = "10.3324/haematol.2014.119537",
language = "English",
volume = "100",
pages = "439--451",
journal = "Haematologica",
issn = "0390-6078",
publisher = "Ferrata Storti Foundation",
number = "4",

}

TY - JOUR

T1 - Role of the clathrin adaptor PICALM in normal hematopoiesis and polycythemia vera pathophysiology

AU - Ishikawa, Yuichi

AU - Maeda, Manami

AU - Pasham, Mithun

AU - Aguet, Francois

AU - Tacheva-Grigorova, Silvia K.

AU - Masuda, Takeshi

AU - Yi, Hai

AU - Lee, Sung Uk

AU - Xu, Jian

AU - Teruya-Feldstein, Julie

AU - Ericsson, Maria

AU - Mullally, Ann

AU - Heuser, John

AU - Kirchhausen, Tom

AU - Maeda, Takahiro

PY - 2015/1/1

Y1 - 2015/1/1

N2 - Clathrin-dependent endocytosis is an essential cellular process shared by all cell types. Despite this, precisely how endocytosis is regulated in a cell-type-specific manner and how this key pathway functions physiologically or pathophysiologically remain largely unknown. PICALM, which encodes the clathrin adaptor protein PICALM, was originally identified as a component of the CALM/AF10 leukemia oncogene. Here we show, by employing a series of conditional Picalm knockout mice, that PICALM critically regulates transferrin uptake in erythroid cells by functioning as a cell-type-specific regulator of transferrin receptor endocytosis. While transferrin receptor is essential for the development of all hematopoietic lineages, Picalm was dispensable for myeloid and B-lymphoid development. Furthermore, global Picalm inactivation in adult mice did not cause gross defects in mouse fitness, except for anemia and a coat color change. Freeze-etch electron microscopy of primary erythroblasts and live-cell imaging of murine embryonic fibroblasts revealed that Picalm function is required for efficient clathrin coat maturation. We showed that the PICALM PIP2 binding domain is necessary for transferrin receptor endocytosis in erythroblasts and absolutely essential for erythroid development from mouse hematopoietic stem/progenitor cells in an erythroid culture system. We further showed that Picalm deletion entirely abrogated the disease phenotype in a Jak2V617F knock-in murine model of polycythemia vera. Our findings provide new insights into the regulation of cell-type-specific transferrin receptor endocytosis in vivo. They also suggest a new strategy to block cellular uptake of transferrin-bound iron, with therapeutic potential for disorders characterized by inappropriate red blood cell production, such as polycythemia vera.

AB - Clathrin-dependent endocytosis is an essential cellular process shared by all cell types. Despite this, precisely how endocytosis is regulated in a cell-type-specific manner and how this key pathway functions physiologically or pathophysiologically remain largely unknown. PICALM, which encodes the clathrin adaptor protein PICALM, was originally identified as a component of the CALM/AF10 leukemia oncogene. Here we show, by employing a series of conditional Picalm knockout mice, that PICALM critically regulates transferrin uptake in erythroid cells by functioning as a cell-type-specific regulator of transferrin receptor endocytosis. While transferrin receptor is essential for the development of all hematopoietic lineages, Picalm was dispensable for myeloid and B-lymphoid development. Furthermore, global Picalm inactivation in adult mice did not cause gross defects in mouse fitness, except for anemia and a coat color change. Freeze-etch electron microscopy of primary erythroblasts and live-cell imaging of murine embryonic fibroblasts revealed that Picalm function is required for efficient clathrin coat maturation. We showed that the PICALM PIP2 binding domain is necessary for transferrin receptor endocytosis in erythroblasts and absolutely essential for erythroid development from mouse hematopoietic stem/progenitor cells in an erythroid culture system. We further showed that Picalm deletion entirely abrogated the disease phenotype in a Jak2V617F knock-in murine model of polycythemia vera. Our findings provide new insights into the regulation of cell-type-specific transferrin receptor endocytosis in vivo. They also suggest a new strategy to block cellular uptake of transferrin-bound iron, with therapeutic potential for disorders characterized by inappropriate red blood cell production, such as polycythemia vera.

UR - http://www.scopus.com/inward/record.url?scp=84926353224&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84926353224&partnerID=8YFLogxK

U2 - 10.3324/haematol.2014.119537

DO - 10.3324/haematol.2014.119537

M3 - Article

C2 - 25552701

AN - SCOPUS:84926353224

VL - 100

SP - 439

EP - 451

JO - Haematologica

JF - Haematologica

SN - 0390-6078

IS - 4

ER -