Intrathecal Infusion of Autologous Adipose-Derived Regenerative Cells in Autoimmune Refractory Epilepsy: Evaluation of Safety and Efficacy

Abstract

Objective/Purpose. Evaluation of efficacy and safety of autologous adipose-derived regenerative cells (ADRCs) treatment in autoimmune refractory epilepsy. Patients. Six patients with proven or probable autoimmune refractory epilepsy (2 with Rasmussen encephalitis, 2 with antineuronal autoantibodies in serum, and 2 with possible FIRES) were included in the project with approval of the Bioethics Committee.

Method: Intrathecal injection of autologous ADRC acquired through liposuction followed by enzymatic isolation was performed. The procedure was repeated 3 times every 3 months with each patient. Neurological status, brain MRI, cognitive function, and antiepileptic effect were monitored during 12 months.

Results: Immediately after the procedure, all patients were in good condition. In some cases, transient mildly elevated body temperature, pain in regions of liposuction, and slight increasing number of seizures during 24 hours were observed. During the next months, some improvements in school, social functioning, and manual performance were observed in all patients. One patient has been seizure free up to the end of trial. In other patients, frequency of seizures was different: from reduced number to the lack of improvement (3-year follow-up).

Conclusion: Autologous ADRC therapy may emerge as a promising option for some patients with autoimmune refractory epilepsy. Based on our trial and other clinical data, the therapy appears to be safe and feasible. Antiepileptic efficacy proved to be various; however, some abilities improved in all children. No signs of psychomotor regression were observed during the first year following the treatment.

Conflict of interest statement

The authors declare that there is no a competing financial interest or conflict of interest regarding the publication of this paper.

Copyright © 2020 Elzbieta Szczepanik et al.

Figures

Scheme 1

Scheme of patients' evaluation. The diagram presents the applied neurological and electrophysiological methods and psychological and biochemical assessment, along with their assignment to time of assessment.

Figure 1

EEG of patient no 3. (a, b) EEG before ARDC treatment (Nov. 2015): spike and spike-slow wave complex: continuous on the left temporal and centro-parieto-occipital region and frequent on the right centro-parietal region. (c) EEG after the last ARDC treatment (Apr. 2016): sharp waves and spikes: frequent on the left temporal and centro-parieto-occipital region and a few on the right centro-parietal region.

Figure 2

Qualitative assessment of cytokines in CSF. (a) Membrane-based antibody arrays for the parallel determination of the relative levels of human cytokines and chemokines from CSF of patients with epilepsy (from left side: P3, P4, and P6). (b) Table—detected and chosen analytes for luminex multiplex cytokine analysis method.

Figure 3

Photos of patient's notebook. (a) Before the first ADRC infusion. (b) 5 months after ADRC infusion—manual ability improvement is visible.

Figure 4

Intrathecal IgG synthesis. Gradual decrease in intrathecal synthesis of IgG after a 3- and 6-month ADRC treatment, followed by an increase in the next 6 months since the last administration; N < 0.01 mg/dl, P1-P6—resp., patient 1–6.

Figure 5

Quantitative analysis of cytokine and chemokine level in CSF during ADRC treatment. Comparison of Angiogenin, IL-10, CXCL10/SDF-1α, Osteopontin, and TNF-α concentration, determined by the Luminex multiplex cytokine analysis method, in CSF, before and after ADRC application (black graph bars/t1—before ADRC application, light grey bars/t2—3 months after 1st ADRC application, medium grey bars/t3—3 months after 2nd ADRC application, dark grey bars/t4—6 months after 3rd ADRC application, P3—respond patient number 3, P6—mild responder, P4—nonresponder.

Figure 6

ADRC in vitro analysis. (a) Flow cytometry analysis—surface markers analysis of ADRC. (b) Quantification of CFU frequency—P3: responder, P4: mild responder, P6: nonresponder, t2: ADRC isolated after 2nd liposuction, t3: ADRC isolated after 3rd liposuction. (c) CFU morphology from P3: (A) paraclone; (B) meroclone-like structures, scale bar—500 μM. (d) The morphology of ADRC derived from P3, scale bar—100 μM. (e) Secretory properties of ADRC exposed to patients CSF: Luminex multiplex cytokine analysis method. Dark grey bars: control, 48 h after incubation with CSF aspirated from responder (P3), mild responder (P6), and nonresponder (P4) before therapy.