Transmembrane Stem Cell Factor Protein Therapeutics Enhance Revascularization in Ischemia without Mast Cell Activation

AbstractStem cell factor (SCF) is a cytokine that regulates hematopoiesis and other biological processes. While clinical treatments using SCF would be highly beneficial, these have been limited by toxicity related to mast cell activation. Transmembrane SCF (tmSCF) has differential activity from soluble SCF and has not been explored as a therapeutic agent. We created novel therapeutics using tmSCF embedded in proteoliposomes or lipid nanodiscs. Mouse models of anaphylaxis and ischemia revealed the tmSCF-based therapies did not activate mast cells and improved the revascularization in the ischemic hind limb. Proteoliposomal tmSCF preferentially acted on endothelial cells to induce angiogenesis while tmSCF nanodiscs had greater activity in inducing stem cell mobilization and recruitment to the site of injury. The type of lipid nanocarrier used altered the relative cellular uptake pathways and signaling in a cell type dependent manner. Overall, we found that tmSCF-based therapies can provide therapeutic benefits without off target effects.

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Abstract 15720: Transmembrane Stem Cell Factor Delivered in Nanocarriers Promotes Angiogenesis in Ischemia Without Mast Cell Activation

Circulation ◽

10.1161/circ.142.suppl_3.15720 ◽

2020 ◽

Vol 142 (Suppl_3) ◽

Author(s):

Eri Takematsu ◽

Sanjana Srinath ◽

Michael Sherman ◽

Andrew K Dunn ◽

Aaron Baker

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Mast Cells ◽

Stem Cell Factor ◽

Cell Activation ◽

The Body ◽

Mast Cell Activation ◽

Protein Therapy ◽

Current Standard ◽

Therapeutic Benefits

Introduction: The current standard cares for peripheral artery disease (PAD) include surgical revascularizations with bypass grafting or percutaneous interventions. However, these interventions cannot be performed in a significant portion of patients, and many do not respond to these surgical procedures. Protein therapy to stimulate the body to create new vasculature is another alternative, which is minimally invasive to patients. Stem cell factor (SCF) is a candidate protein for treating PAD, but clinical use of SCF has been limited due to toxicity related to mast cell activation. SCF also exists in a transmembrane form (tmSCF), possessing differential activities from soluble SCF and has not been explored as a therapeutic agent. Results: To develop tmSCF as a therapeutic we created tmSCF embedded in liposome or lipid nanodisc (Fig. A) . Hindlimb ischemia model on WT and ob/ob mice showed that tmSCF proteliposome (tmSCFPL) and nanodisc (tmSCFND) improved blood flow recovery significantly more than control (Fig. B, C) . Mouse model of anaphylaxis revealed that tmSCF-based therapies did not activate mast cells (Fig. D, E) . Colocalization assay of c-Kit and clathrin/caveolin revealed that mast cells preferentially use clathrin-mediated pathways to internalize SCF and caveolin-mediated pathways for tmSCF-based therapies (Fig. F, G) . Surface c-Kit internalization study on mast cells showed faster uptake of SCF in comparison to tmSCF-based therapies (Fig. H) . Previous study indicates that clathrin-mediated internalization causes increased activation of mast cells. Our studies together with the previous finding suggest that mast cell activation does not occur for tmSCF-based therapies because of the slower uptake, greater utilization of the caveolin internalization pathway and weaker activation of mast cells. Conclusions: TmSCF-based therapies can provide therapeutic benefits without off-target effects on mast cells by tuning activation with nanocarriers.

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The rat c-kit ligand, stem cell factor, induces c-kit receptor-dependent mouse mast cell activation in vivo. Evidence that signaling through the c-kit receptor can induce expression of cellular function.

Journal of Experimental Medicine ◽

10.1084/jem.175.1.245 ◽

1992 ◽

Vol 175 (1) ◽

pp. 245-255 ◽

Cited By ~ 88

Author(s):

B K Wershil ◽

M Tsai ◽

E N Geissler ◽

K M Zsebo ◽

S J Galli

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Mast Cells ◽

Stem Cell Factor ◽

Cell Activation ◽

Mast Cell Activation ◽

Tyrosine Kinase Receptor ◽

Kit Ligand ◽

Kit Receptor

Interactions between products of the mouse W locus, which encodes the c-kit tyrosine kinase receptor, and the Sl locus, which encodes a ligand for c-kit receptor, which we have designated stem cell factor (SCF), have a critical role in the development of mast cells. Mice homozygous for mutations at either locus exhibit several phenotypic abnormalities including a virtual absence of mast cells. Moreover, the c-kit ligand SCF can induce the proliferation and maturation of normal mast cells in vitro or in vivo, and also can result in repair of the mast cell deficiency of Sl/Sld mice in vivo. We now report that administration of SCF intradermally in vivo results in dermal mast cell activation and a mast cell-dependent acute inflammatory response. This effect is c-kit receptor dependent, in that it is not observed when SCF is administered to mice containing dermal mast cells expressing functionally inactive c-kit receptors, is observed with both glycosylated and nonglycosylated forms of SCF, and occurs at doses of SCF at least 10-fold lower on a molar basis than the minimally effective dose of the classical dermal mast cell-activating agent substance P. These findings represent the first demonstration in vivo that a c-kit ligand can result in the functional activation of any cellular lineage expressing the c-kit receptor, and suggest that interactions between the c-kit receptor and its ligand may influence mast cell biology through complex effects on proliferation, maturation, and function.

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Stem Cell Factor Programs the Mast Cell Activation Phenotype

The Journal of Immunology ◽

10.4049/jimmunol.1103366 ◽

2012 ◽

Vol 188 (11) ◽

pp. 5428-5437 ◽

Cited By ~ 61

Author(s):

Tomonobu Ito ◽

Daniel Smrž ◽

Mi-Yeon Jung ◽

Geethani Bandara ◽

Avanti Desai ◽

...

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Stem Cell Factor ◽

Cell Activation ◽

Mast Cell Activation

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Recombinant stem cell factor-induced mast cell activation and smooth muscle contraction in human bronchi.

American Journal of Respiratory Cell and Molecular Biology ◽

10.1165/ajrcmb.11.6.7524570 ◽

1994 ◽

Vol 11 (6) ◽

pp. 646-650 ◽

Cited By ~ 21

Author(s):

B J Undem ◽

L M Lichtenstein ◽

W C Hubbard ◽

S Meeker ◽

J L Ellis

Keyword(s):

Mast Cell ◽

Smooth Muscle ◽

Stem Cell ◽

Muscle Contraction ◽

Stem Cell Factor ◽

Cell Activation ◽

Smooth Muscle Contraction ◽

Mast Cell Activation ◽

Human Bronchi

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gp49B1 suppresses stem cell factor-induced mast cell activation-secretion and attendant inflammation in vivo

European Journal of Immunology ◽

10.1002/eji.200323978 ◽

2003 ◽

Vol 33 (8) ◽

pp. 2262-2268 ◽

Cited By ~ 22

Author(s):

Anna M. Feldweg ◽

Daniel S. Friend ◽

Joseph S. Zhou ◽

Yoshihide Kanaoka ◽

Massoud Daheshia ◽

...

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Stem Cell Factor ◽

Cell Activation ◽

Mast Cell Activation

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Calcineurin–Rcan1 Interaction Contributes to Stem Cell Factor–Mediated Mast Cell Activation

The Journal of Immunology ◽

10.4049/jimmunol.1301271 ◽

2013 ◽

Vol 191 (12) ◽

pp. 5885-5894 ◽

Cited By ~ 12

Author(s):

Zhengli Wu ◽

Yanhong Li ◽

Adam J. MacNeil ◽

Robert D. Junkins ◽

Jason N. Berman ◽

...

Keyword(s):

Mast Cell ◽

Stem Cell ◽

Stem Cell Factor ◽

Cell Activation ◽

Mast Cell Activation

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Lysophosphatidic acid induces human mast cell activation in an IL-4-dependent manner*1

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2003.12.277 ◽

2004 ◽

Vol 113 (2) ◽

pp. S83

Author(s):

D LIN

Keyword(s):

Mast Cell ◽

Lysophosphatidic Acid ◽

Cell Activation ◽

Mast Cell Activation ◽

Human Mast Cell ◽

Dependent Manner

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Quercetin Attenuates Trauma-Induced Heterotopic Ossification by Tuning Immune Cell Infiltration and Related Inflammatory Insult

Frontiers in Immunology ◽

10.3389/fimmu.2021.649285 ◽

2021 ◽

Vol 12 ◽

Author(s):

Juehong Li ◽

Ziyang Sun ◽

Gang Luo ◽

Shuo Wang ◽

Haomin Cui ◽

...

Keyword(s):

Mast Cell ◽

Heterotopic Ossification ◽

Cell Activation ◽

Immune Cell ◽

Inflammatory Responses ◽

Macrophage Polarization ◽

Mast Cell Activation ◽

Cell Infiltration ◽

Immune Cell Infiltration ◽

Dependent Manner

Heterotopic ossification (HO) is one of the most intractable disorders following musculoskeletal injury and is characterized by the ectopic presence of bone tissue in the soft tissue leading to severe loss of function in the extremities. Recent studies have indicated that immune cell infiltration and inflammation are involved in aberrant bone formation. In this study, we found increased monocyte/macrophage and mast cell accumulation during early HO progression. Macrophage depletion by clodronate liposomes and mast cell stabilization by cromolyn sodium significantly impeded HO formation. Therefore, we proposed that the dietary phytochemical quercetin could also suppress immune cell recruitment and related inflammatory responses to prevent HO. As expected, quercetin inhibited the monocyte-to-macrophage transition, macrophage polarization, and mast cell activation in vitro in a dose-dependent manner. Using a murine burn/tenotomy model, we also demonstrated that quercetin attenuated inflammatory responses and HO in vivo. Furthermore, elevated SIRT1 and decreased acetylated NFκB p65 expression were responsible for the mechanism of quercetin, and the beneficial effects of quercetin were reversed by the SIRT1 antagonist EX527 and mimicked by the SIRT agonist SRT1720. The findings in this study suggest that targeting monocyte/macrophage and mast cell activities may represent an attractive approach for therapeutic intervention of HO and that quercetin may serve as a promising therapeutic candidate for the treatment of trauma-induced HO by modulating SIRT1/NFκB signaling.

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Recent advances in mast cell activation and regulation

F1000Research ◽

10.12688/f1000research.22037.1 ◽

2020 ◽

Vol 9 ◽

pp. 196 ◽

Cited By ~ 4

Author(s):

Hwan Soo Kim ◽

Yu Kawakami ◽

Kazumi Kasakura ◽

Toshiaki Kawakami

Keyword(s):

Mast Cell ◽

Mast Cells ◽

Cell Activation ◽

De Novo ◽

Immunoglobulin E ◽

Secretory Granules ◽

Structural Features ◽

Mast Cell Activation ◽

Dependent Manner ◽

Drug Induced

Mast cells are innate immune cells that intersect with the adaptive immunity and play a crucial role in the initiation of allergic reactions and the host defense against certain parasites and venoms. When activated in an allergen- and immunoglobulin E (IgE)-dependent manner, these cells secrete a large variety of allergenic mediators that are pre-stored in secretory granules or de novo–synthesized. Traditionally, studies have predominantly focused on understanding this mechanism of mast cell activation and regulation. Along this line of study, recent studies have shed light on what structural features are required for allergens and how IgE, particularly anaphylactic IgE, is produced. However, the last few years have seen a flurry of new studies on IgE-independent mast cell activation, particularly via Mrgprb2 (mouse) and MRGPRX2 (human). These studies have greatly advanced our understanding of how mast cells exert non-histaminergic itch, pain, and drug-induced pseudoallergy by interacting with sensory neurons. Recent studies have also characterized mast cell activation and regulation by interleukin-33 (IL-33) and other cytokines and by non-coding RNAs. These newly identified mechanisms for mast cell activation and regulation will further stimulate the allergy/immunology community to develop novel therapeutic strategies for treatment of allergic and non-allergic diseases.

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