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Sensory neuropathy hampers nociception-mediated bone marrow stem cell release in mice and patients with diabetes.

Dang Z, Maselli D, Spinetti G, Sangalli E, Carnelli F, Rosa F, Seganfreddo E, Canal F, Furlan A, Paccagnella A, Paiola E, Lorusso B, Specchia C, Albiero M, Cappellari R, Avogaro A, Falco A, Quaini F, Ou K, Rodriguez-Arabaolaza I, Emanueli C, Sambataro M, Fadini GP, Madeddu P - Diabetologia (2015)

Bottom Line: Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation.Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.Nociceptors may represent a new target for treatment of diabetic complications.

View Article: PubMed Central - PubMed

Affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, Upper Maudlin Street, Bristol, BS2 8HW, UK.

ABSTRACT

Aims/hypothesis: Upon tissue injury, peripheral sensory neurons release nociceptive factors (e.g. substance P [SP]), which exert local and systemic actions including the recruitment of bone marrow (BM)-derived haematopoietic stem and progenitor cells (HSPCs) endowed with paracrine pro-angiogenic properties. We herein explore whether diabetic neuropathy interferes with these phenomena.

Methods: We first investigated the presence of sensory neuropathy in the BM of patients with type 2 diabetes by immunohistochemistry and morphometry analyses of nerve size and density and assessment of SP release by ELISA. We next analysed the association of sensory neuropathy with altered HSPC release under ischaemia or following direct stimulation with granulocyte colony-stimulating factor (G-CSF). BM and circulating HSPCs expressing the neurokinin 1 receptor (NK1R), which is the main SP receptor, were measured by flow cytometry. We finally assessed whether an altered modulation of SP secretion interferes with the mobilisation and homing of NK1R-HSPCs in a mouse model of type 2 diabetes after limb ischaemia (LI).

Results: Nociceptive fibres were reduced in the BM of patients and mice with type 2 diabetes. Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation. Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.

Conclusions/interpretation: Sensory neuropathy translates into defective liberation and homing of reparative HSPCs. Nociceptors may represent a new target for treatment of diabetic complications.

No MeSH data available.


Related in: MedlinePlus

Impaired release of CD34+NK1R+ cells in patients with type 2 diabetes and neurovascular complications. (a) Immunofluorescence micrograph of BM CD34+NK1R+ cells (scale bar, 20 μm). ND, non-diabetic. Arrowheads show cells positive for CD34 and NK1R markers. (b–d) Bar graphs showing the levels of CD34+ (b) and CD34+NK1R+ cells (c) and relative abundance of NK1R+ cells over total CD34+ cells (d). Black bars, non-diabetic (n = 6); white bars, T2DM-U (n = 3); light grey bars, T2DM-N (n = 5); dark grey, T2DM-NI (n = 10). (e–g) Bar graphs showing the abundance of CD34+ (e) and NK1R+ cells on total CD34+ cells (f) (p = 0.06, non-diabetic vs T2DM-NI) in the PB of non-diabetic individuals and patients with type 2 diabetes, assessed by flow cytometry (non-diabetic, n = 8; T2DM-U, n = 5; T2DM-N, n = 12; T2DM-NI, n = 11); the NK1R mean fluorescence intensity on CD34+ cells is shown in (g). (h) Dot plots of gating strategy. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs non-diabetic controls
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Fig3: Impaired release of CD34+NK1R+ cells in patients with type 2 diabetes and neurovascular complications. (a) Immunofluorescence micrograph of BM CD34+NK1R+ cells (scale bar, 20 μm). ND, non-diabetic. Arrowheads show cells positive for CD34 and NK1R markers. (b–d) Bar graphs showing the levels of CD34+ (b) and CD34+NK1R+ cells (c) and relative abundance of NK1R+ cells over total CD34+ cells (d). Black bars, non-diabetic (n = 6); white bars, T2DM-U (n = 3); light grey bars, T2DM-N (n = 5); dark grey, T2DM-NI (n = 10). (e–g) Bar graphs showing the abundance of CD34+ (e) and NK1R+ cells on total CD34+ cells (f) (p = 0.06, non-diabetic vs T2DM-NI) in the PB of non-diabetic individuals and patients with type 2 diabetes, assessed by flow cytometry (non-diabetic, n = 8; T2DM-U, n = 5; T2DM-N, n = 12; T2DM-NI, n = 11); the NK1R mean fluorescence intensity on CD34+ cells is shown in (g). (h) Dot plots of gating strategy. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs non-diabetic controls

Mentions: Having documented the presence of structural and functional nociceptive alterations in the BM of patients with type 2 diabetes, we next conducted immunofluorescence and flow cytometry analyses of CD34+ HSPCs, which co-express the SP receptor NK1R. Patients with complicated diabetes showed a depletion of CD34+NK1R+ HSPCs in their BM (Fig. 3a–d) and PB (Fig. 3e–h). Mean fluorescence intensity analysis indicated that NK1R was downregulated in cells from diabetic patients (Fig. 3g). Furthermore, superimposed CLI failed to induce an increment in circulating CD34+NK1R+ HSPCs (Fig. 3e–h). Altogether, these data indicate that sensory neuropathy is associated with BM depletion and altered release of a subpopulation of HSPCs that is responsive to SP chemoattraction. In particular, the presence of ischaemia, which reportedly acts as a potent stimulus for CD34+NK1R+ HSPC release [4], fails to promote cell mobilisation in patients with type 2 diabetes.Fig. 3


Sensory neuropathy hampers nociception-mediated bone marrow stem cell release in mice and patients with diabetes.

Dang Z, Maselli D, Spinetti G, Sangalli E, Carnelli F, Rosa F, Seganfreddo E, Canal F, Furlan A, Paccagnella A, Paiola E, Lorusso B, Specchia C, Albiero M, Cappellari R, Avogaro A, Falco A, Quaini F, Ou K, Rodriguez-Arabaolaza I, Emanueli C, Sambataro M, Fadini GP, Madeddu P - Diabetologia (2015)

Impaired release of CD34+NK1R+ cells in patients with type 2 diabetes and neurovascular complications. (a) Immunofluorescence micrograph of BM CD34+NK1R+ cells (scale bar, 20 μm). ND, non-diabetic. Arrowheads show cells positive for CD34 and NK1R markers. (b–d) Bar graphs showing the levels of CD34+ (b) and CD34+NK1R+ cells (c) and relative abundance of NK1R+ cells over total CD34+ cells (d). Black bars, non-diabetic (n = 6); white bars, T2DM-U (n = 3); light grey bars, T2DM-N (n = 5); dark grey, T2DM-NI (n = 10). (e–g) Bar graphs showing the abundance of CD34+ (e) and NK1R+ cells on total CD34+ cells (f) (p = 0.06, non-diabetic vs T2DM-NI) in the PB of non-diabetic individuals and patients with type 2 diabetes, assessed by flow cytometry (non-diabetic, n = 8; T2DM-U, n = 5; T2DM-N, n = 12; T2DM-NI, n = 11); the NK1R mean fluorescence intensity on CD34+ cells is shown in (g). (h) Dot plots of gating strategy. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs non-diabetic controls
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Related In: Results  -  Collection

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Fig3: Impaired release of CD34+NK1R+ cells in patients with type 2 diabetes and neurovascular complications. (a) Immunofluorescence micrograph of BM CD34+NK1R+ cells (scale bar, 20 μm). ND, non-diabetic. Arrowheads show cells positive for CD34 and NK1R markers. (b–d) Bar graphs showing the levels of CD34+ (b) and CD34+NK1R+ cells (c) and relative abundance of NK1R+ cells over total CD34+ cells (d). Black bars, non-diabetic (n = 6); white bars, T2DM-U (n = 3); light grey bars, T2DM-N (n = 5); dark grey, T2DM-NI (n = 10). (e–g) Bar graphs showing the abundance of CD34+ (e) and NK1R+ cells on total CD34+ cells (f) (p = 0.06, non-diabetic vs T2DM-NI) in the PB of non-diabetic individuals and patients with type 2 diabetes, assessed by flow cytometry (non-diabetic, n = 8; T2DM-U, n = 5; T2DM-N, n = 12; T2DM-NI, n = 11); the NK1R mean fluorescence intensity on CD34+ cells is shown in (g). (h) Dot plots of gating strategy. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 vs non-diabetic controls
Mentions: Having documented the presence of structural and functional nociceptive alterations in the BM of patients with type 2 diabetes, we next conducted immunofluorescence and flow cytometry analyses of CD34+ HSPCs, which co-express the SP receptor NK1R. Patients with complicated diabetes showed a depletion of CD34+NK1R+ HSPCs in their BM (Fig. 3a–d) and PB (Fig. 3e–h). Mean fluorescence intensity analysis indicated that NK1R was downregulated in cells from diabetic patients (Fig. 3g). Furthermore, superimposed CLI failed to induce an increment in circulating CD34+NK1R+ HSPCs (Fig. 3e–h). Altogether, these data indicate that sensory neuropathy is associated with BM depletion and altered release of a subpopulation of HSPCs that is responsive to SP chemoattraction. In particular, the presence of ischaemia, which reportedly acts as a potent stimulus for CD34+NK1R+ HSPC release [4], fails to promote cell mobilisation in patients with type 2 diabetes.Fig. 3

Bottom Line: Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation.Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.Nociceptors may represent a new target for treatment of diabetic complications.

View Article: PubMed Central - PubMed

Affiliation: Bristol Heart Institute, School of Clinical Sciences, University of Bristol, Upper Maudlin Street, Bristol, BS2 8HW, UK.

ABSTRACT

Aims/hypothesis: Upon tissue injury, peripheral sensory neurons release nociceptive factors (e.g. substance P [SP]), which exert local and systemic actions including the recruitment of bone marrow (BM)-derived haematopoietic stem and progenitor cells (HSPCs) endowed with paracrine pro-angiogenic properties. We herein explore whether diabetic neuropathy interferes with these phenomena.

Methods: We first investigated the presence of sensory neuropathy in the BM of patients with type 2 diabetes by immunohistochemistry and morphometry analyses of nerve size and density and assessment of SP release by ELISA. We next analysed the association of sensory neuropathy with altered HSPC release under ischaemia or following direct stimulation with granulocyte colony-stimulating factor (G-CSF). BM and circulating HSPCs expressing the neurokinin 1 receptor (NK1R), which is the main SP receptor, were measured by flow cytometry. We finally assessed whether an altered modulation of SP secretion interferes with the mobilisation and homing of NK1R-HSPCs in a mouse model of type 2 diabetes after limb ischaemia (LI).

Results: Nociceptive fibres were reduced in the BM of patients and mice with type 2 diabetes. Patients with neuropathy showed a remarkable reduction in NK1R-HSPC mobilisation under ischaemia or upon G-CSF stimulation. Following LI, diabetic mice manifested an altered SP gradient between BM, peripheral blood and limb muscles, accompanied by a depressed recruitment of NK1R-HSPCs to the ischaemic site.

Conclusions/interpretation: Sensory neuropathy translates into defective liberation and homing of reparative HSPCs. Nociceptors may represent a new target for treatment of diabetic complications.

No MeSH data available.


Related in: MedlinePlus