Regulation of adult hematopoiesis by the a disintegrin and metalloproteinase 10 (ADAM10)

doi:10.1016/j.bbrc.2013.11.020

Biochemical and Biophysical Research Communications

Volume 442, Issues 3–4, 13 December 2013, Pages 234-241

https://doi.org/10.1016/j.bbrc.2013.11.020 Get rights and content

Highlights

•
ADAM10 deficiency leads to the development of a myeloproliferative disorder.
•
Bone marrow transplantation experiments strongly support a non-cell autonomous origin.
•
A central role of ADAM10 for the regulation of granulopoiesis and lymphopoiesis is suggested.

Abstract

Adult hematopoiesis requires tightly regulated cell–cell interactions between hematopoietic cells and the bone marrow stromal microenvironment. We addressed the question if the ectodomain sheddase ADAM10 is essential to regulate adult hematopoiesis. Induced ADAM10 deletion in hematopoietic cells resulted in morphological and histological abnormalities that resemble an unclassified myeloproliferative disorder (MPD). The MPD was characterized by an expansion of granulocytic subpopulations and their infiltration of peripheral hematopoietic tissues, the development of hepatosplenomegaly with extramedullary erythropoiesis, lymphnodepathy and death of the mice around 20 weeks after induction. ADAM10 expression analysis during the different stages of the MPD revealed that non-targeted hematopoietic cells repopulated the immune system of the ADAM10-deficient mice. Examination of mice with a myeloid- or epidermis-specific deletion of ADAM10 and bone marrow transplantation (BMT) experiments indicated that the development of the MPD can be triggered by non-cell autonomous effects. We found that plasma levels of clinical markers for MPD such as G-CSF, TIMP-1 and IL-16 were significantly elevated in ADAM10-deficient mice. Our findings indicate that a tightly controlled ADAM10 expression is needed to balance hematopoietic cell-fate decisions in adult mice.

Introduction

In recent studies emerging evidence for the crucial role of tightly controlled cell–cell communication between the bone marrow microenvironment and hematopoietic cells during hematopoiesis has been established. However the detailed molecular mechanisms regulating this process have remained elusive so far. A rapid and irreversible posttranslational modification of cell surface protein density can be achieved by an extracellular proteolytic cleavage step which is referred to as “ectodomain shedding” [1], [2], [3], [4]. ADAM10 is an ubiquitously expressed protein which is capable of cleaving numerous transmembrane proteins such as Notch receptors [5] that are implicated for proper regulation of hematopoietic development and homeostasis. In order to study the consequence of ADAM10 downregulation during adult hematopoiesis, we conditionally inactivated its expression using the inducible Mx-Cre strain [6] and a myeloid-specific deleter strain [7], respectively. The Mx promoter becomes activated by interferon response or synthetic double-stranded RNA in most cells of immunologically important tissues including bone marrow stroma cells [6]. A similarly induced knockout of the Notch1 receptor and the putative sole downstream signaling mediator Rbp-j, led to de novo ectopic B-cell development in the thymus from common lymphoid progenitors [8] thus underlining the importance of Notch receptor signaling for cell-fate decisions during lymphopoiesis.

In this study, similar to a recently published report [9], we show that the induced loss of ADAM10 leads to the development and progression of a MPD with concomitant loss of mature B- and T-lymphocytes in the bone marrow, spleen and peripheral blood. The expanded myeloid population mainly consists of CD11b⁺/Gr1⁺ cells with normal ADAM10 expression levels. Additionally bone marrow transplantation (BMT) experiments strongly support a non-cell autonomous origin of the MPD which is possibly triggered and maintained by the excessive secretion of G-CSF, TIMP-1 and IL-16.

Section snippets

Generation of inducible conditional ADAM10 (cKO) mice

The ADAM10^flox/flox mice were crossed with Mx-Cre mice [10], K5-tTA-Cre mice [11] or LysM-Cre mice [7]. To induce Mx-mediated deletion of ADAM10 during adult hematopoiesis, 6-week old ADAM10^flox/flox/Mx-Cre^+/− (ADAM10^−/−) and ADAM10^flox/flox (Ctr) mice were injected three times in two-day intervals with 250 μg polyinosinic–polycytidylic acid (pI–pC).

Western blot of tissue lysates

Tissues were uniformly lysed (150 mM NaCl, 2 mM EDTA, 0.8 mM EGTA, 10 mM Tris–HCl, pH 7.4 and Complete Protease Inhibitors (1:25, Roche)). ADAM10 was

Lack of ADAM10 expression and activity in the adult hematopoietic system

We induced the deletion of the protease in ADAM10^{flox/flox; Mx1Cre} mice and sacrificed mice 2 weeks after the last injection of double stranded RNA (pI–pC). Genomic deletion was observed in bone marrow, spleen, liver and skin (Fig. 1A). Immunoblot analysis revealed a strong downregulation of ADAM10 protein expression in bone marrow and spleen as well as of the proform of the protease in liver (Fig. 1B). Despite significantly downregulated ADAM10 protein expression (Fig. 1CI) in bone marrow

Discussion and conclusion

Since the metalloproteinase ADAM10 is a well-known control point to modulate cell–cell contacts [13], [14] we aimed to analyze if this protease plays a cell autonomous or a non-cell autonomous role in the hematopoiesis of adult mice. With the generation of conditional ADAM10-deficient mice under the control of the interferon-inducible Mx-promoter we were able to address this question. The different time points chosen for our analysis resemble the chronic and acceleration phases of MPD patients.

Acknowledgments

This work was supported by the Deutsche Forschungsgemeinschaft SFB877. M.D. is supported by the Netherlands Heart Foundation (Dr. E. Dekker post-doctoral fellow Grant 2007T034 and 2012T079).

References (31)

K. Reiss et al.
Breaking up the tie: disintegrin-like metalloproteinases as regulators of cell migration in inflammation and invasion
Pharmacol. Ther.
(2006)
R.M. Wang et al.
Effect of rare earth on the microstructures and properties of a low expansion superalloy
Micron
(2002)
P. Sinha et al.
Myeloid-derived suppressor cells express the death receptor Fas and apoptose in response to T cell-expressed FasL
Blood
(2011)
M. Yoda et al.
Dual functions of cell-autonomous and non-cell-autonomous ADAM10 activity in granulopoiesis
Blood
(2011)
I. Diamond et al.
Conditional gene expression in the epidermis of transgenic mice using the tetracycline-regulated transactivators tTA and rTA linked to the keratin 5 promoter
J. Invest. Dermatol.
(2000)
A. Hikita et al.
Negative regulation of osteoclastogenesis by ectodomain shedding of receptor activator of NF-kappaB ligand
J. Biol. Chem.
(2006)
M.R. Zocchi et al.
High ERp5/ADAM10 expression in lymph node microenvironment and impaired NKG2D-ligands recognition in Hodgkin lymphomas
Blood
(2012)
L. Zhou et al.
Notch-dependent control of myelopoiesis is regulated by fucosylation
Blood
(2008)
Y.W. Kim et al.
Defective Notch activation in microenvironment leads to myeloproliferative disease
Blood
(2008)
A. Amour et al.
The in vitro activity of ADAM-10 is inhibited by TIMP-1 and TIMP-3
FEBS Lett.
(2000)

T. Maretzky et al.

ADAM10-mediated E-cadherin release is regulated by proinflammatory cytokines and modulates keratinocyte cohesion in eczematous dermatitis

J. Invest. Dermatol.

(2008)

K. Tanigaki et al.

Regulation of lymphocyte development by Notch signaling

Nat. Immunol.

(2007)

H. Han et al.

Inducible gene knockout of transcription factor recombination signal binding protein-J reveals its essential role in T versus B lineage decision

Int. Immunol.

(2002)

D. Hartmann et al.

The disintegrin/metalloprotease ADAM 10 is essential for Notch signalling but not for alpha-secretase activity in fibroblasts

Hum. Mol. Genet.

(2002)

B.E. Clausen et al.

Conditional gene targeting in macrophages and granulocytes using LysMcre mice

Transgenic Res.

(1999)

Cited by (14)

ADAM10 and Neprilysin level decreases in immune cells of mice bearing metastatic breast carcinoma: Possible role in cancer inflammatory response
2024, International Immunopharmacology
ADAM10 and Neprilysin, proteases, play critical role in inflammatory disease, however their role in cancer immune response is not clear. We here evaluated changes in immune response using an experimental model for breast cancer.
Highly metastatic breast cancer cells (4T1-derived) were injected orthotopically (mammary-pad of Balb-c mice) to induce tumors. Changes in enzyme level and activity as well as alterations in inflammatory cytokine release in the presence or absence of ADAM10 and NEP activity was determined using specific inhibitors and recombinant proteins. Cytokine response was evaluated using mix leucocyte cultures obtained from control and tumor-bearing mice. ANOVA with Dunnett's posttest was used for statistical analysis.
ADAM10 and NEP expression was decreased markedly in lymph nodes and spleens of tumor-bearing mice. ADAM10 activity was reduced together with apparent alterations of ADAM10 processing. ADAM10 and NEP activity decreased TNF-α, IL-6 and IFN-ɣ secretion. Suppression of these inflammatory cytokines were more prominent in cultures obtained from control mice demonstrating counteracting factors that are exist in tumor-bearing mice.
Loss of ADAM10 and NEP activity in immune cells during breast cancer metastasis might be one of the main factors involved in induction of chronic inflammation by tumors.
The extracellular matrix of hematopoietic stem cell niches
2022, Advanced Drug Delivery Reviews
Citation Excerpt :
Clinical markers for myeloproliferative disorder such as elevated TIMP-1 levels in blood plasma were observed in these mice. This study strongly suggest that ADAM-10 is necessary for a balanced myeloid/lymphoid cell-fate decision of HSPCs [438]. The four mammalian tissue inhibitors of metalloproteinases (TIMP1-4), originally identified as collagenase inhibitors, are secreted proteins that can inhibit all activated MMPs [423,439].
Hematopoietic stem cells (HSCs) are the life-long source of all types of blood cells. Their function is controlled by their direct microenvironment, the HSC niche in the bone marrow. Although the importance of the extracellular matrix (ECM) in the niche by orchestrating niche architecture and cellular function is widely acknowledged, it is still underexplored. In this review, we provide a comprehensive overview of the ECM in HSC niches. For this purpose, we first briefly outline HSC niche biology and then review the role of the different classes of ECM molecules in the niche one by one and how they are perceived by cells. Matrix remodeling and the emerging importance of biophysics in HSC niche function are discussed. Finally, the application of the current knowledge of ECM in the niche in form of artificial HSC niches for HSC expansion or targeted differentiation as well as drug testing is reviewed.
Role of ADAM10 in intestinal crypt homeostasis and tumorigenesis
2017, Biochimica et Biophysica Acta - Molecular Cell Research
Citation Excerpt :
Subsequently, NEXT then undergoes γ -secretase dependent processing to generate NICD. Numerous studies in mice have confirmed that ADAM10 is the principal sheddase responsible for ligand-dependent Notch S2 cleavage in vivo [58–66]. In addition, in vitro studies have established that canonical ligand-induced proteolysis of human NOTCH-1, -2 and-3 receptors is strictly dependent upon ADAM10 [67].
A disintegrin and metalloproteinases (ADAMs) are a family of mSultidomain, membrane-anchored proteases that regulate diverse cellular functions, including cell adhesion, migration, proteolysis and other cell signaling events. Catalytically-active ADAMs act as ectodomain sheddases that proteolytically cleave type I and type II transmembrane proteins and some GPI-anchored proteins from the cellular surface. ADAMs can also modulate other cellular signaling events through a process known as regulated intramembrane proteolysis (RIP). Through their proteolytic activity, ADAMs can rapidly modulate key cell signaling pathways in response to changes in the extracellular environment (e.g. inflammation) and play a central role in coordinating intercellular communication. Dysregulation of these processes through aberrant expression, or sustained ADAM activity, is linked to chronic inflammation, inflammation-associated cancer and tumorigenesis. ADAM10 was the first disintegrin-metalloproteinase demonstrated to have proteolytic activity and is the prototypic ADAM associated with RIP activity (e.g. sequential Notch receptor processing). ADAM10 is abundantly expressed throughout the gastrointestinal tract and during normal intestinal homeostasis ADAM10 regulates many cellular processes associated with intestinal development, cell fate specification and maintenance of intestinal stem cell/progenitor populations. In addition, several signaling pathways that undergo ectodomain shedding by ADAM10 (e.g. Notch, EGFR/ErbB, IL-6/sIL-6R) help control intestinal injury/regenerative responses and may drive intestinal inflammation and colon cancer initiation and progression. Here, I review some of the proposed functions of ADAM10 associated with intestinal crypt homeostasis and tumorigenesis within the gastrointestinal tract in vivo. This article is part of a Special Issue entitled: Proteolysis as a Regulatory Event in Pathophysiology edited by Stefan Rose-John.
Leukocytes require ADAM10 but not ADAM17 for their migration and inflammatory recruitment into the alveolar space
2014, Blood
Citation Excerpt :
Thus, ADAM10 deficiency leads to accumulation of slow-migrating cells in the chemotaxis membranes and interstitial accumulation within the lung. Interestingly, Mx-driven ADAM10 deficiency in mice leads to leukocyte accumulation in the spleen,27,28 and this effect could be in part due to the inability of leukocytes to pass through the spleen filter. Additionally, B cell-specific ADAM10 deficiency results in impaired marginal zone development and asthmatic responses,29,30 which might also depend on ADAM10-mediated B-cell migration.
Inflammation is a key process in various diseases, characterized by leukocyte recruitment to the inflammatory site. This study investigates the role of a disintegrin and a metalloproteinase (ADAM) 10 and ADAM17 for leukocyte migration in vitro and in a murine model of acute pulmonary inflammation. Inhibition experiments or RNA knockdown indicated that monocytic THP-1 cells and primary human neutrophils require ADAM10 but not ADAM17 for efficient chemokine-induced cell migration. Signaling and adhesion events that are linked to cell migration such as p38 and ρ GTPase-family activation, F-actin polymerization, adhesion to fibronectin, and up-regulation of α₅ integrin were also dependent on ADAM10 but not ADAM17. This was confirmed with leukocytes isolated from mice lacking either ADAM10 or ADAM17 in all hematopoietic cells (vav 1 guanine nucleotide exchange factor [Vav]-Adam10^−/− or Vav-Adam17^−/− mice). In lipopolysaccharide-induced acute pulmonary inflammation, alveolar recruitment of neutrophils and monocytes was transiently increased in Vav-Adam17^−/− but steadily reduced in Vav-Adam10^−/− mice. This deficit in alveolar leukocyte recruitment was also observed in LysM-Adam10^−/− mice lacking ADAM10 in myeloid cells and correlated with protection against edema formation. Thus, with regard to leukocyte migration, leukocyte-expressed ADAM10 but not ADAM17 displays proinflammatory activities and may therefore serve as a target to limit inflammatory cell recruitment.
Targeting cardiomyocyte ADAM10 ectodomain shedding promotes survival early after myocardial infarction
2022, Nature Communications
Host T Cell Dedifferentiation Effects Drive HIV-1 Latency Stability
2022, Journal of Virology

View all citing articles on Scopus

¹: Present address: Heart Research Centre Göttingen, Universitätsmedizin Göttingen, Department of Cardiology and Pneumology, University Göttingen, Göttingen, Germany.

View full text

Regulation of adult hematopoiesis by the a disintegrin and metalloproteinase 10 (ADAM10)

Highlights

Abstract

Introduction

Section snippets

Generation of inducible conditional ADAM10 (cKO) mice

Western blot of tissue lysates

Lack of ADAM10 expression and activity in the adult hematopoietic system

Discussion and conclusion

Acknowledgments

Pharmacol. Ther.

Micron

Blood

Blood

J. Invest. Dermatol.

J. Biol. Chem.

Blood

Blood

Blood

FEBS Lett.

J. Invest. Dermatol.

Regulation of lymphocyte development by Notch signaling

Nat. Immunol.

Inducible gene knockout of transcription factor recombination signal binding protein-J reveals its essential role in T versus B lineage decision

Int. Immunol.

The disintegrin/metalloprotease ADAM 10 is essential for Notch signalling but not for alpha-secretase activity in fibroblasts

Hum. Mol. Genet.

Conditional gene targeting in macrophages and granulocytes using LysMcre mice

Transgenic Res.