Humoral Influence of Repeated Lineage-Negative Stem/Progenitor Cell Administration on Articulatory Functions in ALS Patients

Abstract

Amyotrophic lateral sclerosis (ALS) remains a fatal, neurodegenerative disease frequently leading to dysarthria and impaired swallowing. Better understanding of ALS pathophysiology is prompting the use of humoral cell therapies. Hence, a repeated cellular therapy was applied to ALS patients as an attempt to prevent speech deterioration. Autologous bone marrow-derived lineage-negative (Lin^-) cells were intrathecally administered three times at six-week intervals to 42 sporadic ALS patients. Patients were examined for articulatory functions using subjective (VHI) and objective (FDA) scales. Selected trophic, proinflammatory factors and expression profiles of miRNA were measured in cerebrospinal fluid (CSF) and plasma by multiplex Luminex and q-PCR in different timepoints. Of the 42 patients who received the Lin^- cells, 6 showed improvement in articulatory functions, 27 remained stable, and 9 deteriorated after 18 weeks of therapy according to FDA scale. Clinical improvement was particularly evident by the 7^th day of each cell application and concerned better cough and swallow reflex, soft palate, laryngeal time, pitch, and volume. These results correlated with significant changes in the concentration of various trophic and proinflammatory factors and miRNA expression profiles. A multiple application of Lin^- cells proved to be safe and feasible. The repeated procedure can potentate a humoral effect and prevent speech deterioration. A short-lasting trophic effect of each Lin^- cells administration was observed on local and systemic level. However, further in-depth studies are necessary to sustain the beneficial effect.

Figure 1

Functional assessment results with ALS-FRSr (a) and Norris scale (b) in three groups of sALS patients in time. Green—group I; black—group II; red—group III. The results are presented as the mean values. Squares represent the analyzed timepoints.

Figure 2

FDA scale assessment results after 7 and 28 days post each injection of Lin⁻ cells (No. 1, No. 2, No. 3) compared to the baseline values (day 0 of each procedure). Colors represent the percentage of patients (n =42) in which the analyzed parameter improved (green), remained stable (gray) or deteriorated (red).

Figure 3

Subjective VHI questionnaire results after 7 and 28 days post each injection of Lin⁻ cells (No. 1, No. 2, and No. 3) compared to the baseline values (day 0 of each procedure). Colors represent the percentage of patients (n = 42) in which the analyzed parameter improved (green), remained stable (gray), or deteriorated (red).

Figure 4

Concentrations of analyzed factors in CSF (pg/mL) of 42 sALS patients divided into three groups, based on the FDA scale results. The samples were collected in six timepoints presented on the x-axis. Roman numerals represent each Lin⁻ cell administration (I, II, and III), and Arabic numbers stand for the day of sample collection (0 and 7). Data are presented as the median values with an interquartile range. Level of significance: ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. BDNF: brain-derived neurotrophic factor; NGF: nerve growth factor; NT-3: neurotrophin 3; CRP: C-reactive protein.

Figure 5

Concentrations of analyzed factors in plasma (pg/mL) of 42 sALS patients divided into three groups, based on the FDA scale results. The samples were collected in nine timepoints presented on the x-axis. Roman numerals represent each Lin⁻ cell administration (I, II, and III), and Arabic numbers stand for the day of sample collection (0, 7, and 28). Data are presented as the median values with an interquartile range. Level of significance: ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001. BDNF: brain-derived neurotrophic factor; NGF: nerve growth factor; NT-3: neurotrophin 3; CRP: C-reactive protein.

Figure 6

Results of qPCR miRNA expression analysis in (a) CSF and (b) plasma assessed in 42 sALS patients divided into three groups, based on the FDA scale results. The samples were collected in six timepoints presented on the x-axis. Roman numerals represent each Lin⁻ cell administration (I, II, and III), and Arabic numbers stand for the day of sample collection (0 and 7). Data are presented as the median values with an interquartile range. Level of significance: ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001.