case_images: PMC11438342_01
case_images: Schematic of overall procedures.. (A) Nerve from a non-amyotrophic lateral sclerosis donor is used to derive a Schwann cell culture (B) and then generate allogeneic extracellular vesicles released by Schwann cells through media ultracentrifugation, which are then assayed for effects on neuronal culture (C). Exosomes are then infused and circulated with potential effects on central nervous system inflammation and trophic/supportive effects in muscles and nerves, including at the neuromuscular junction. Blood is sampled from the patient for biomarkers, and clinical readouts are performed.
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case_images: NRR.NRR-D-23-01815-F1
case_images: When cultured with allogeneic control SCEVs generated from normal (younger, non-ALS) donor SCs (Figure 2B), an enhancement in the patient's SC vitality was observed, evidenced by an increased expression of the SC marker (CD 271) at 38.51% (Figure 1D), as compared to 20.43% (Figure 2C) without the non-ALS SCEVs. The pre-infusion rate of functional decline (ALSFRS-R) (Mandrioli et al., 2015) shown in Figure 3 averaged 1.71 points/month, followed by seven measurements during the infusion period with an average rate of decrease of 1.22 points per month (difference 0.49 points per month).
case_images: Patient Schwann cell culture analyses.. Schwann cell cultures from the ALS subject with and without added allogenic extracellular vesicles released by Schwann cells (SCEV) from a donor without ALS. Phase contrast microscope, original magnification, 10x. Without (A) or with allogeneic SCEV (B) was added. (C, D) Flow cytometric analysis for Schwann cell marker CD271, and fibroblast marker CD90, same conditions as A and B. In D, with allogeneic SCEV, the percent of CD271 is increased and that of CD90 is decreased.
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case_images: The patient's SCs appeared dysfunctional (they exhibited a rounded morphology with stunted processes) and proliferated poorly (Figure 2A), they could not be expanded beyond passage 2. The cultured SCs were interspersed with numerous fibroblasts (Figure 2A), affecting the purity of the culture. When cultured with allogeneic control SCEVs generated from normal (younger, non-ALS) donor SCs (Figure 2B), an enhancement in the patient's SC vitality was observed, evidenced by an increased expression of the SC marker (CD 271) at 38.51% (Figure 1D), as compared to 20.43% (Figure 2C) without the non-ALS SCEVs. In addition, the presence of fibroblasts (CD90) was reduced to 29.10% (Figure 2D) as compared to 50.65% (Figure 2C).
case_images: Patient's forced vital capacity and ALSFRS-R.. Values of FVC and ALSFRS-R during the study. The pre-infusion period was 70 days. The period in which the seven infusions occurred is indicated on the x-axis with the red bar, and with the pink boxes. There were two values after the infusions were paused. The stabilization of values is indicated by the pink boxes. Overall slopes are shown for the pre-infusion and infusion periods, including all values. ALSFRS-R: Amyotrophic Lateral Sclerosis Revised Functional Rating Scale; FVC: forced vital capacity.
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case_images: PMC11438342_01_NRR-20-1207-g003.jpg
case_images: NRR.NRR-D-23-01815-F3
case_images: The pre-infusion rate of functional decline (ALSFRS-R) (Mandrioli et al., 2015) shown in Figure 3 averaged 1.71 points/month, followed by seven measurements during the infusion period with an average rate of decrease of 1.22 points per month (difference 0.49 points per month). The shaded boxes (Figure 3) illustrate an apparent stabilization of the disease during the infusion period. Pre-infusion pulmonary function declined as expected in ALS (Schiffman and Belsh, 1993; forced vital capacity in liters measured by spirometry), as shown in Figure 3 upper panel.