Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative condition characterized by rapid degeneration of motoneurons (MNs), leading to progressive muscle atrophy and, ultimately, mortality within a few years of diagnosis. Although the precise mechanisms initiating MN degeneration are not fully understood, the involvement of non-neuronal cells, including microglia, in ALS pathophysiology is increasingly recognized. Mesenchymal stromal cell (MSC)-based therapies have emerged as a promising avenue for ALS treatment, yet clinical outcomes remain variable, underscoring the necessity for additional pre-clinical investigations. This study evaluated the therapeutic potential of human MSCs derived from Wharton's jelly (WJMSC) in the female SOD1G93A mouse model of ALS. Our results indicated that intravenous administration of WJMSC during the presymptomatic phase of the disease notably delayed the onset of motor deficits and extended the lifespan. This functional benefit was associated with the preservation of MNs in the cervical spinal cord. In the lumbar spinal cord, we did not observe MN neuroprotection, but we noted a temporary increase in microgliosis following WJMSC treatment. Our results supported the therapeutic benefits of human MSC in ALS, while also highlighting the differential responses of spinal-cord regions to the treatment during the disease progression. This study underscores the importance of targeting specific disease stages and regions for MSC therapy in ALS, paving the way for refined and potentially more effective therapeutic strategies.

Introduction: Diabetes mellitus (DM) has become a public health problem, which is associated with high morbidity and mortality, due to the chronic complications, such as diabetic neuropathy. Current recommendations for the treatment of neuropathic pain achieve a reduction of 30% in only 30% of cases. Therefore, it is necessary to identify new therapeutic approaches to improve the quality of life of diabetic patients. Methods: This work evaluated the antinociceptive effect of intranasal administration of the combination of dextro-ketamine (keta), a non-competitive glutamatergic receptor antagonist, and dexmedetomidine (DEX), a selective alpha2-adrenergic agonist, in rats with neuropathic pain induced by streptozotocin-DM. Results: The thermal hyperalgesia and mechanical allodynia observed in DM model are reduced with the intranasal administration of the combination of keta and DEX (200 + 0.10 mu g/kg) after 3 days of treatment. The antinociceptive action could be due to reduction of Ca2+influx with lower glutamate release and reduced excitability through the activation of alpha2-adrenergic receptors by DEX and reduction of NMDA receptor activation by glutamate with lower excitability due to the antagonism produced by keta. DM induced increased expression of glial fibrillary acid protein (GFAP) and tumor necrosis factor-alpha (TNF-alpha) detected by immunohistochemistry, indicating greater astrocyte activity and intense inflammatory response. Intranasal administration for 10 days of the combination of low doses of keta and DEX promoted an intense decrease in the expression of both GFAP and TNF-alpha, indicating lower activation of astrocytes in the spinal cord and reduced production and release of TNF-alpha, favoring the reduction of inflammation. Conclusion: Intranasal administration of low doses of keta with DEX could be a new therapeutic approach to reduce neuropathic pain and consequently improve the quality of life of diabetic patients.

For most of the 20th century, the dogma was that new neurons could not be generated in the adult mammalian brain. Nowadays, neurogenesis in the mammalian is widely accepted, especially in two brain regions: the ventricular-subventricular zone (V-SVZ) around the lateral ventricles and the subgranular zone (SGZ) in the hippocampus. The cytoarchitecture of these neurogenic niches has been widely studied, and different cell types and molecules have been identified, but the permanence of neural stem cells versus progenitor cells in these niches remains elusive. Neurogenesis may be modulated by physiological and pathological processes. Understanding the signaling pathways that modulate neurogenesis is a potential therapeutic strategy to avoid its decline associated with aging and/or pathology, as well as to replace lost cells after injury or degenerative diseases. Lastly, neural stem cells isolated from neurogenic regions or generated from pluripotent stem cells are also promising future therapeutic strategies.

Diabetes mellitus-related morbidity and mortality are primarily caused by long-term complications such as retinopathy, nephropathy, cardiomyopathy, and neuropathy. Diabetic neuropathy (DN) involves the progressive degeneration of axons and nerve fibers due to chronic exposure to hyperglycemia. This metabolic disturbance leads to excessive activation of the glycolytic pathway, inducing oxidative stress and mitochondrial dysfunction, ultimately resulting in nerve damage. There is no specific treatment for painful DN, and new approaches should aim not only to relieve pain but also to prevent oxidative stress and reduce inflammation. Given that existing therapies for painful DN are not effective for diabetic patients, mesenchymal stromal cells (MSCs)-based therapy shows promise for providing immunomodulatory and paracrine regulatory functions. MSCs from various sources can improve neuronal dysfunction associated with DN. Transplantation of MSCs has led to a reduction in hyperalgesia and allodynia, along with the recovery of nerve function in diabetic rats. While the pathogenesis of diabetic neuropathic pain is complex, clinical trials have demonstrated the importance of MSCs in modulating the immune response in diabetic patients. MSCs reduce the levels of inflammatory factors and increase anti-inflammatory cytokines, thereby interfering with the progression of DM. Further investigation is necessary to ensure the safety and efficacy of MSCs in preventing or treating neuropathic pain in diabetic patients.

The umbilical cord is a perinatal tissue normally discarded after delivery that can be used as a source of different cell types with promising therapeutic potential. In this chapter, we discuss the use of different cell populations isolated from umbilical cord tissue and blood, which include hematopoietic stem cells/progenitors, mononuclear cells, and mesenchymal stem/stromal cells, as therapeutic agents for neurological disorders. We review the main preclinical studies and clinical trials based on umbilical cord derived cells in optic neuropathies and retinal diseases, stroke, and neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis.

Chronic pain presents a major challenge in contemporary medicine, given the limited effectiveness and numerous adverse effects linked to available treatments. Recognizing the potential of the cholinergic pathway as a therapeutic target, the present work evaluates the antinociceptive activity of a combination of Cris-104, a novel α4β2* receptor agonist, and donepezil, a central anticholinesterase agent. Isobolographic analysis revealed that equimolar combination was approximately 10 times more potent than theoretically calculated equipotent additive dose. Administration of Cris-104 and donepezil combination (3 μmol/kg) successfully reversed hyperalgesia and mechanical allodynia observed in rats subjected to spinal nerve ligation (SNL). The combination also modulated neuroinflammation by reducing astrocyte activation, evident in the decreased expression of glial fibrillary acidic protein (GFAP) in the spinal cord. The observed synergism in combining a nicotinic receptor agonist with an anticholinesterase agent underscores its potential for treating chronic pain. This alternative therapeutic distinct advantage, including dose reduction and high selectivity for the receptor, contribute to a more favorable profile with minimized adverse effects.

EDITORIAL article Front. Cell. Neurosci., 29 September 2023Sec. Non-Neuronal Cells Volume 17 - 2023 | https://doi.org/10.3389/fncel.2023.1287049

Gangliosides, sialic acid-containing sphingolipids, are major constituents of neuronal membranes. According to the number of sialic acids and the structure of the oligosaccharide chain, gangliosides can be classified as simple or complex and grouped in different ganglio-series. Hundreds of gangliosides have been identified in vertebrate cells, with different expression patterns during development and related to several physiological processes, especially in the nervous system. While GD3 and its O-acetylated form, 9acGD3, are highly expressed in early developmental stages, GM1, GD1a, GD1b, and GT1b are the most abundant ganglioside species in the mature nervous system. Mutations in enzymes involved in ganglioside metabolism can lead to the accumulation of specific species, a condition termed gangliosidosis and usually marked by severe neurological impairment. Changes in ganglioside levels have also been described in several neurodegenerative diseases, such as Alzheimer's and Parkinson's. In this review, we summarized recent information about the roles of GD3, 9acGD3, GM1, GD1a, GD1b, GT1b, and other ganglioside species in nervous system development and regeneration, as well as clinical trials evaluating possible therapeutic applications of these molecules.

Alzheimer's disease is a severe, highly disabling neurodegenerative disease, clinically characterized by a progressive decline in cognitive functions, and is the most common form of dementia in the elderly. For decades, the search for disease-modifying therapies has focused on the two main Alzheimer's disease histopathological hallmarks, seeking to prevent, mitigate, or clear the formation of extracellular aggregates of β-amyloid peptide and intracellular neurofibrillary tangles of tau protein, although without clinical success. Mesenchymal stem cell-based therapy has emerged as a promising alternative for the treatment of Alzheimer's disease, especially because it also targets other crucial players in the pathogenesis of the disease, such as neuroinflammation, synaptic dysfunction/loss, oxidative stress, and impaired neurogenesis. Herein, we review current knowledge of the therapeutic potential of mesenchymal stem cells and their extracellular vesicles for Alzheimer's disease, discussing the most recent findings in both preclinical and clinical trials as well as how advanced technologies have helped to overcome some limitations and contributed to stimulate the development of more effective treatments.

Neurological disorders include a wide spectrum of clinical conditions affecting the central and peripheral nervous systems. For these conditions, which affect hundreds of millions of people worldwide, generally limited or no treatments are available, and cell-based therapies have been intensively investigated in preclinical and clinical studies. Among the available cell types, mesenchymal stem/stromal cells (MSCs) have been widely studied but as yet no cell-based treatment exists for neurological disease. We review current knowledge of the therapeutic potential of MSC-based therapies for neurological diseases, as well as possible mechanisms of action that may be explored to hasten the development of new and effective treatments. We also discuss the challenges for culture conditions, quality control, and the development of potency tests, aiming to generate more efficient cell therapy products for neurological disorders.

Induced pluripotent stem cells (iPSCs) are generated from adult cells that have been reprogrammed to pluripotency. However, in vitro cultivation and genetic reprogramming increase genetic instability, which could result in chromosomal abnormalities. Maintenance of genetic stability after reprogramming is required for possible experimental and clinical applications. The aim of this study was to analyze chromosomal alterations by using the G-banding karyotyping method applied to 97 samples from 38 iPSC cell lines generated from peripheral blood or Wharton's jelly. Samples from patients with long QT syndrome, Jervell and Lange-Nielsen syndrome and amyotrophic lateral sclerosis and from normal individuals revealed the following chromosomal alterations: acentric fragments, chromosomal fusions, premature centromere divisions, double minutes, radial figures, ring chromosomes, polyploidies, inversions and trisomies. An analysis of two samples generated from Wharton's jelly before and after reprogramming showed that abnormal clones can emerge or be selected and generate an altered lineage. IPSC lines may show clonal and nonclonal chromosomal aberrations in several passages (from P6 to P34), but these aberrations are more common in later passages. Many important chromosomal aberrations were detected, showing that G-banding is very useful for evaluating genetic instability with important repercussions for the application of iPSC lines.

ABSTRACT  Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease which leads to a progressive degeneration of motoneurons. Since the pharmacological options available provide only a slight increase in life expectancy, cell therapy is emerging as a promising therapeutic alternative for ALS. A growing body of evidence from studies using genetically engineered ALS animal models demonstrate the safety and efficacy of therapies based on different cell types such as mononuclear cells, neural progenitors, and mesenchymal stem cells. Despite the encouraging results in preclinical studies, cell therapy-based clinical trials for ALS have achieved only modest results so far, probably due to the genotypic variations seen among ALS patients, which is difficult to reproduce in animal models. The advent of induced pluripotent stem cells (iPSCs) has enabled the development of patient-specific cell lines, a valuable tool to investigate in vitro molecular mechanisms of the disease and therapies in different genetic backgrounds. The applications of ALS iPSCs and their future therapeutic potential are also briefly discussed in this chapter.

Stem cell therapy is a promising alternative approach to heart diseases. The most prevalent source of multipotent stem cells, usually called somatic or adult stem cells (mesenchymal stromal/stem cells, MSCs) used in clinical trials is bone marrow (BM-MSCs), adipose tissue (AT-MSCs), umbilical cord (UC-MSCs) and placenta. Therapeutic use of MSCs in cardiovascular diseases is based on the benefits in reducing cardiac fibrosis and inflammation that compose the cardiac remodeling responsible for the maintenance of normal function, something which may end up causing progressive and irreversible dysfunction. Many factors lead to cardiac fibrosis and failure, and an effective therapy is lacking to reverse or attenuate this condition. Different approaches have been shown to be promising in surpassing the poor survival of transplanted cells in cardiac tissue to provide cardioprotection and prevent cardiac remodeling. This review includes the description of pre-clinical and clinical investigation of the therapeutic potential of MSCs in improving ventricular dysfunction consequent to diverse cardiac diseases.

Background Anthracyclines (AC) are chemotherapeutic drugs used for treatment of various types of cancer. However, up to 26% of patients develop anthracycline-induced cardiotoxicity (AIC), ranging from mild arrhythmias to severe ventricular dysfunction. This study investigates the ability of human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) to recapitulate in vitro the susceptibility that certain patients have to develop AIC. Methods Patients treated with cumulative dose ≥200 mg/m2 of AC were recruited, as well as age- and gender-matched healthy volunteers (HV). iPSCs were derived from erythroblasts, evaluated for pluripotency and genomic stability, and differentiated into CM. At day 30, CMs were treated for 72h with 0.01-100µM Doxorubicin (DOX) alone or in presence of 1-100µM cardioprotectant Dexrazoxane (DRZ), 1-100µM of its metabolite ADR925 or 1mM N-acetylcysteine (NAC). Treated CMs were evaluated for viability and sarcomeric organization. Next-Generation Sequencing (NGS) was performed for cardiac targets in iPSC (5 AC-sensitive, 5 AC-resistant). Results iPSCs presented normal karyotype (G-banding assay) and pluripotency (RT-PCR: Oct4, Sox2, Nanog, Klf4; flow cytometry: OCT4, SOX2, NANOG; spontaneous differentiation into three germ layers RT-PCR: Tubulinβ3, Bmp4, Afp). CMs expressed cardiac Troponin T (cTnT) (AC-S: 82.45±14.09%; HV: 90.45±4.99%; n=6). Increasing doses of DOX led to higher mortality rates (AC-S: 0.1µM–15.09±6.33%; 1µM–46.62±13.27%; 10µM–64.46±21.99%; 100µM–96.1±5.55%; n=10; HV: 0.1µM–3.65±16.37%; 1µM–27.35±25.56%; 10µM–56.2±25.82%; 100µM–89.75±11.63%; n=6). Cotreatment with cardioprotectants showed no significant difference in cell viability (IC50 in µM AC-S: DOX=3.18±2.95, n=10; DOX+100µM DRZ=2.84±2.24, n=5, p=0.82; DOX+100µM ADR925=5.56±2.91, n=4, p=0.19; DOX+1mM NAC=2.45±2.51, n=6, p=0.62; HV: DOX=10.23±10.07, n=6; DOX+100µM DRZ=1.39, n=1; DOX+100µM ADR925=6.56±7.02, n=3, p=0.59; DOX+1mM NAC=3.64±4.72, n=4, p=0.26). cTnT staining revealed sarcomeric disorganization for DOX≥10µM. NGS sequencing revealed 49 nonsynonymous variants, none of which associated with AIC. Only one of those variants, located at desmoplakin gene, was present in all 4 AC-S patients and absent in AC-R ones. Conclusion iPSC from AC-S and AC-R patients were successfully generated and differentiated into CM. Viability tests recapitulated the patient's clinical cardiotoxicity and confirmed iPSC-CM as a platform for drug screening. Anthracyclines (AC) are chemotherapeutic drugs used for treatment of various types of cancer. However, up to 26% of patients develop anthracycline-induced cardiotoxicity (AIC), ranging from mild arrhythmias to severe ventricular dysfunction. This study investigates the ability of human-induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) to recapitulate in vitro the susceptibility that certain patients have to develop AIC. Patients treated with cumulative dose ≥200 mg/m2 of AC were recruited, as well as age- and gender-matched healthy volunteers (HV). iPSCs were derived from erythroblasts, evaluated for pluripotency and genomic stability, and differentiated into CM. At day 30, CMs were treated for 72h with 0.01-100µM Doxorubicin (DOX) alone or in presence of 1-100µM cardioprotectant Dexrazoxane (DRZ), 1-100µM of its metabolite ADR925 or 1mM N-acetylcysteine (NAC). Treated CMs were evaluated for viability and sarcomeric organization. Next-Generation Sequencing (NGS) was performed for cardiac targets in iPSC (5 AC-sensitive, 5 AC-resistant). iPSCs presented normal karyotype (G-banding assay) and pluripotency (RT-PCR: Oct4, Sox2, Nanog, Klf4; flow cytometry: OCT4, SOX2, NANOG; spontaneous differentiation into three germ layers RT-PCR: Tubulinβ3, Bmp4, Afp). CMs expressed cardiac Troponin T (cTnT) (AC-S: 82.45±14.09%; HV: 90.45±4.99%; n=6). Increasing doses of DOX led to higher mortality rates (AC-S: 0.1µM–15.09±6.33%; 1µM–46.62±13.27%; 10µM–64.46±21.99%; 100µM–96.1±5.55%; n=10; HV: 0.1µM–3.65±16.37%; 1µM–27.35±25.56%; 10µM–56.2±25.82%; 100µM–89.75±11.63%; n=6). Cotreatment with cardioprotectants showed no significant difference in cell viability (IC50 in µM AC-S: DOX=3.18±2.95, n=10; DOX+100µM DRZ=2.84±2.24, n=5, p=0.82; DOX+100µM ADR925=5.56±2.91, n=4, p=0.19; DOX+1mM NAC=2.45±2.51, n=6, p=0.62; HV: DOX=10.23±10.07, n=6; DOX+100µM DRZ=1.39, n=1; DOX+100µM ADR925=6.56±7.02, n=3, p=0.59; DOX+1mM NAC=3.64±4.72, n=4, p=0.26). cTnT staining revealed sarcomeric disorganization for DOX≥10µM. NGS sequencing revealed 49 nonsynonymous variants, none of which associated with AIC. Only one of those variants, located at desmoplakin gene, was present in all 4 AC-S patients and absent in AC-R ones. iPSC from AC-S and AC-R patients were successfully generated and differentiated into CM. Viability tests recapitulated the patient's clinical cardiotoxicity and confirmed iPSC-CM as a platform for drug screening.

Introduction: Amyotrophic Lateral Sclerosis (ALS) is an adult-onset progressive and fatal neurodegenerative disease that selectively affects upper and lower motor neurons. Death occurs within 3 to 5 years of onset, usually from respiratory complications. Most cases of ALS are sporadic (SALS), but familial forms of the disease (FALS) represent approximately 10% of the cases. More than 30 genes have been associated with ALS and mutations in these genes account for more than a half of all familial cases and about 10% of sporadic cases. One of the most prevalent genes is TARDBP, responsible for approximately 4-6% of FALS and nearly 1-2% of SALS cases. The aim of this study was to perform the screening of known ALS genes, to increase the knowledge of the mutations that circulate in the population from Rio de Janeiro. Methods: The screening of mutations was performed through the Illumina Next Generation Sequencing (NGS) platform with the use of a sequencing panel that contained the TARDBP, SOD1, FUS, VAPB, SMN1 and SMN2 genes. Results: A novel missense mutation (p.Phe194Leu) in exon 5 of the TARDBP gene was found in a sporadic male patient who died at the age of 58 (2018). The mutation, a TTT/CTT substitution, was not detected in any mutation databases and in the literature. In silico analysis of this variant with different algorithms were performed and the results pointed to a probably damaging impact and that the mutation is disease causing. Conclusion: Through the study of the ALS genes by the NGS, we were able to identify a novel TARDBP mutation in a non-familial ALS patient. In addition, this study also increases the number of known TARDBP mutations in ALS patients and our knowledge of the mutations that affect the patients from of population from Rio de Janeiro.

Gangliosides are glycosphingolipids abundantly expressed in the vertebrate nervous system, and are classified into a-, b-, or c-series according to the number of sialic acid residues. The enzyme GD3 synthase converts GM3 (an a-series ganglioside) into GD3, a b-series ganglioside highly expressed in the developing and adult retina. The present study evaluated the visual system of GD3 synthase knockout mice (GD3s(-/-)), morphologically and functionally. The absence of b- series gangliosides in the retinas of knockout animals was confirmed by mass spectrometry imaging, which also indicated an accumulation of a-series gangliosides, such as GM3. Retinal ganglion cell (RGC) density was significantly reduced in GD3s(-/-) mice, with a similar reduction in the number of axons in the optic nerve. Knockout animals also showed a 15% reduction in the number of photoreceptor nuclei, but no difference in the bipolar cells. The area occupied by GFAP-positive glial cells was smaller in GD3s(-/-) retinas, but the number of microglial cells/macrophages did not change. In addition to the morphological alterations, a 30% reduction in light responsiveness was detected through quantification of pS6-expressing RGC, an indicator of neural activity. Furthermore, electroretinography (ERG) indicated a significant reduction in RGC and photoreceptor electrical activity in GD3s(-/-) mice, as indicated by scotopic ERG and pattern ERG (PERG) amplitudes. Finally, evaluation of the optomotor response demonstrated that GD3s(-/-) mice have reduced visual acuity and contrast sensitivity. These results suggest that b-series gangliosides play a critical role in regulating the structure and function of the mouse visual system.