Dipeptidyl Peptidase-4 Inhibitors for the Potential Treatment of Brain Disorders; A Mini-Review With Special Focus on Linagliptin and Stroke

Cerebral stroke is a leading cause of death and persistent disability of elderly in the world. Although stroke prevention by targeting several risk factors such as diabetes and hypertension has decreased the stroke incidence, the total number of strokes is increasing due to the population aging and new preventive therapies are needed. Moreover, post-stroke acute pharmacological strategies aimed to reduce stroke-induced brain injury have failed in clinical trials despite being effective in animal models. Finally, approximately 30% of surviving stroke patients do not recover from stroke and remain permanently dependent on supportive care in activities of daily living. Therefore, strategies to improve stroke recovery in the post-acute phase are highly needed. Linagliptin is a dipeptidyl peptidase-4 inhibitor which is clinically approved to reduce hyperglycemia in type 2 diabetes. The regulation of glycemia by dipeptidyl peptidase-4 inhibition is mainly achieved by preventing endogenous glucagon-like peptide-1 (GLP-1) degradation. Interestingly, linagliptin has also shown glycaemia-independent beneficial effects in animal models of stroke, Parkinson's disease and Alzheimer's disease. In some case the preclinical data have been supported with some clinical data. Although potentially very interesting for the development of new strategies against stroke and neurodegenerative disorders, the mode of action of linagliptin in the brain is still largely unknown and seems to occur in a GLP-1R-independent manner. The purpose of this mini-review is to summarize and discuss the recent experimental and clinical work regarding the effects of linagliptin in the central nervous system, with special emphasis on acute neuroprotection, stroke prevention and post-stroke recovery. We also highlight the main questions in this research field that need to be addressed in clinical perspective

Glucagon-like peptide-1 receptor activation reduces ischaemic brain damage following stroke in Type 2 diabetic rats

Diabetes is a strong risk factor for premature and severe stroke. The GLP-1R (glucagon-like peptide-1 receptor) agonist Ex-4 (exendin-4) is a drug for the treatment of T2D (Type 2 diabetes) that may also have neuroprotective effects. The aim of the present study was to determine the efficacy of Ex-4 against stroke in diabetes by using a diabetic animal model, a drug administration paradigm and a dose that mimics a diabetic patient on Ex-4 therapy. Furthermore, we investigated inflammation and neurogenesis as potential cellular mechanisms underlying the Ex-4 efficacy. A total of seven 9-month-old Type 2 diabetic Goto–Kakizaki rats were treated peripherally for 4 weeks with Ex-4 at 0.1, 1 or 5 μg/kg of body weight before inducing stroke by transient middle cerebral artery occlusion and for 2–4 weeks thereafter. The severity of ischaemic damage was measured by evaluation of stroke volume and by stereological counting of neurons in the striatum and cortex. We also quantitatively evaluated stroke-induced inflammation, stem cell proliferation and neurogenesis. We show a profound anti-stroke efficacy of the clinical dose of Ex-4 in diabetic rats, an arrested microglia infiltration and an increase of stroke-induced neural stem cell proliferation and neuroblast formation, while stroke-induced neurogenesis was not affected by Ex-4. The results show a pronounced anti-stroke, neuroprotective and anti-inflammatory effect of peripheral and chronic Ex-4 treatment in middle-aged diabetic animals in a preclinical setting that has the potential to mimic the clinical treatment. Our results should provide strong impetus to further investigate GLP-1R agonists for their neuroprotective action in diabetes, and for their possible use as anti-stroke medication in non-diabetic conditions

Neurogenesis and transplantation in stroke-damaged brain

Stroke in young adult rodents triggers neurogenesis in the damaged striatum and intact hippocampus. as stroke happens frequently in aged individuals it is of great importance to know whether aging affects brain recovery mechanisms. Aged and young adult rats were subjected to stroke by transient occlusion of the middle cerebral artery and proliferating cells were labeled by intraperitoneal administration of the mitotic marker BrdU. Animals were sacrificed at 7 weeks after infarct and new cells were examined for expression of BrdU and neuronal and glial markers with epifluorescent and confocal microscopy. Young and aged rats showed similarly increased number of new neurons in the striatum, although basal proliferation was reduced in the aged subventricular zone. In contrast, both basal proliferation and the generation of new neurons was significantly reduced in aged subgranular zone and granule cell layer of the hippocampus. This study shows that basal neurogenesis is impaired in the aged rat brain compared to young, but the brain responds to damage with increased neurogenesis. This increase was similar in the striatum of both young and old animals, indicating the existence of potential self-repair mechanisms in the aged brain. Stem cell transplantation is considered one of the future therapeutic methods for treatment of stroke. Therefore identification and characterization of viable neural stem cell lines is essential for the development of successful therapies. Neural stem cells (NSCs) isolated from fetal human striatum and cortex were studied and compared for their neurogenic capacity in vitro and after transplantation in neonatal intact or adult, stroke-damaged brain. Cortex- and striatum-derived NSCs expanded as neurospheres did not differ in proliferative capacity, growth rate, secondary sphere formation, and expression of general neural markers. However, whereas cortical NSCs produced higher number of glutamatergic and tyrosine hydroxylase- and calretinin-positive neurons, several-fold more neurons expressing the striatal projection neuron marker, DARPP-32, were observed in cultures of striatal NSCs. Human cortical and striatal NSCs survived and migrated equally well after transplantation in neonatal rats. The two NSC types also generated similar numbers of mature NeuN+ neurons, which were several-fold higher at 4 months as compared to at 1 month after grafting. At 4 months, the grafts contained cells with morphological characteristics of neurons, astrocytes, and oligodendrocytes, the majority of neurons expressing parvalbumin. Striatal and cortical NSCs exhibited similar robust survival (30%) at 1 month after transplantation in stroke-subjected animals and migrated throughout the damaged striatum. Striatal NSCs migrated longer distance and occupied a bigger volume of striatum. In the transplantation core, cells were undifferentiated, virtually all expressing cellular markers of immature neural lineage such as nestin, and to lesser extent also GFAP, ?III-tubulin, DCX and calretinin. Immunocytochemistry with proliferation markers (p-H3 and Ki67) revealed that grafted striatal and cortical NSCs cease to proliferate. Human cells outside the transplantation core differentiated, exhibited mature neuronal morphology and expressed adult neuronal markers such as HuD, calbindin and parvalbumin. Interestingly, striatal NSCs generate greater number of parvalbumin+ and calbindin+ neurons and virtually none of the grafted cells differentiated into astrocytes or oligodendrocytes. Based on these data, human fetal striatum- and cortex-derived NSCs could be considered safe and viable sources with strong neurogenic potential for further exploration in animal models of stroke

Neural Stem Cell-Based Therapy for Ischemic Stroke

Stem cell-based approaches for the treatment of stroke have been the subject of intensive research over the past decade. Based on accumulated experimental evidence, stem cell-based therapy is a very promising prospect for the development of a novel treatment to restore stroke-damaged brain and impaired neurological function. Studies performed on experimental animal models of stroke employed a variety of stem cell types from diverse sources and have demonstrated their ability to replace lost neurons and functionally integrate into the brain, modulate inflammation, and stimulate angiogenesis and neurogenesis from an endogenous stem cell pool, most likely through trophic actions. A few clinical trials in stroke patients using stem cell transplantation have been completed or are on-going but the results have not yet proven the effectiveness of the stem cell-based approaches. A joint effort of stroke researchers and clinicians is needed to further optimize treatment protocols using safe and reproducible stem cell sources tested in relevant animal models of stroke and showing substantial neurological recovery of stroke-impaired function

Survival, migration and neuronal differentiation of human fetal striatal and cortical neural stem cells grafted in stroke-damaged rat striatum

Stroke is a neurodegenerative disorder and the leading cause of disability in adult humans. Treatments to support efficient recovery in stroke patients are lacking. Several studies have demonstrated the ability of grafted neural stem cells (NSCs) to partly improve impaired neurological functions in stroke-subjected animals. Recently, we reported that NSCs from human fetal striatum and cortex exhibit region-specific differentiation in vitro, but survive, migrate and form neurons to a similar extent after intrastriatal transplantation in newborn rats. Here, we have transplanted the same cells into the stroke-damaged striatum of adult rats. The two types of NSCs exhibited a similar robust survival (30%) at 1 month after transplantation, and migrated throughout the damaged striatum. Striatal NSCs migrated farther and occupied a larger volume of striatum. In the transplantation core, cells were undifferentiated and expressed nestin and, to a lesser extent, also GFAP, beta III-tubulin, DCX and calretinin, markers of immature neural lineage. Immunocytochemistry using markers of proliferation (p-H3 and Ki67) revealed a very low content of proliferating cells (< 1%) in the grafts. Human cells outside the transplantation core differentiated, exhibited mature neuronal morphology and expressed mature neuronal markers such as HuD, calbindin and parvalbumin. Interestingly, striatal NSCs generated a greater number of parvalbumin(+) and calbindin(+) neurons. Virtually none of the grafted cells differentiated into astrocytes or oligodendrocytes. Based on these data, human fetal striatum- and cortex-derived NSCs could be considered potentially safe and viable for transplantation, with strong neurogenic potential, for further exploration in animal models of stroke

Human fetal cortical and striatal neural stem cells generate region-specific neurons in vitro and differentiate extensively to neurons after intrastriatal transplantation in neonatal rats.

Human fetal brain is a potential source of neural stem cells (NSCs) for cell replacement therapy in neurodegenerative diseases. We explored whether NSCs isolated from cortex and striatum of human fetuses, aged 6-9 weeks post-conception, maintain their regional identity and differentiate into specific neuron types in culture and after intrastriatal transplantation in neonatal rats. We observed no differences between cortex- and striatum-derived NSCs expanded as neurospheres in proliferative capacity, growth rate, secondary sphere formation, and expression of neural markers. After 4 weeks of differentiation in vitro, cortical and striatal NSCs gave rise to similar numbers of GABAergic and VMAT2- and parvalbumin-containing neurons. However, whereas cortical NSCs produced higher number of glutamatergic and tyrosine hydroxylase- and calretinin-positive neurons, several-fold more neurons expressing the striatal projection neuron marker, DARPP-32, were observed in cultures of striatal NSCs. Human cortical and striatal NSCs survived and migrated equally well after transplantation. The two NSC types also generated similar numbers of mature NeuN-positive neurons, which were several-fold higher at 4 months as compared to at 1 month after grafting. At 4 months, the grafts contained cells with morphologic characteristics of neurons, astrocytes, and oligodendrocytes. Many of neurons were expressing parvalbumin. Our data show that NSCs derived from human fetal cortex and striatum exhibit region-specific differentiation in vitro, and survive, migrate, and form mature neurons to the same extent after intrastriatal transplantation in newborn rats