A pilot open-label trial of minocycline in patients with autism and regressive features

Carlos A Pardo, Ashura Buckley, Audrey Thurm, Li-Ching Lee, Arun Azhagiri, David M Neville, Susan E Swedo, Carlos A Pardo, Ashura Buckley, Audrey Thurm, Li-Ching Lee, Arun Azhagiri, David M Neville, Susan E Swedo

Abstract

Background: Minocycline is a tetracycline derivative that readily crosses the blood brain barrier and appears to have beneficial effects on neuroinflammation, microglial activation and neuroprotection in a variety of neurological disorders. Both microglial activation and neuroinflammation have been reported to be associated with autism. The study was designed to evaluate the effects of minocycline treatment on markers of neuroinflammation and autism symptomatology in children with autism and a history of developmental regression.

Methods: Eleven children were enrolled in an open-label trial of six months of minocycline (1.4 mg/kg). Ten children completed the trial. Behavioral measures were collected and cerebrospinal fluid (CSF), serum and plasma were obtained before and at the end of minocycline treatment and were analyzed for markers of neuroinflammation.

Results: Clinical improvements were negligible. The laboratory assays demonstrated significant changes in the expression profile of the truncated form of brain derived neurotrophic factor (BDNF) (P = 0.042) and hepatic growth factor (HGF) (P = 0.028) in CSF. In serum, the ratio of the truncated BDNF form and α-2 macroglobulin (α-2 M), was also significantly lower (P = 0.028) while the mature BDNF/α-2 M ratio revealed no difference following treatment. Only the chemokine CXCL8 (IL-8) was significantly different (P = 0.047) in serum while no significant changes were observed in CSF or serum in chemokines such as CCL2 (MCP-1) or cytokines such as TNF-α, CD40L, IL-6, IFN-γ and IL-1β when pre- and post-treatment levels of these proteins were compared. No significant pre- and post-treatment changes were seen in the profiles of plasma metalloproteinases, putative targets of the effects of minocycline.

Conclusions: Changes in the pre- and post-treatment profiles of BDNF in CSF and blood, HGF in CSF and CXCL8 (IL-8) in serum, suggest that minocycline may have effects in the CNS by modulating the production of neurotrophic growth factors. However, in this small group of children, no clinical improvements were observed during or after the six months of minocycline administration.

Trial registration: NCT00409747.

Keywords: Autism; BDNF; Chemokines; Clinical trial; Cytokines; Metalloproteinases; Microglia; Minocycline; Neuroinflammation; Neurotrophins.

Figures

Figure 1
Figure 1
CXCL8 (IL-8) Post-treatment trend in serum. (A) Serum levels of CXCL8 (IL-8) were significantly lower after treatment with minocycline. (P = 0.047). (B) The trend profile of CXCL8 (IL-8) in serum showed a lowering effect of treatment in seven of ten patients (red lines) while only three of ten (green lines) showed an increase.
Figure 2
Figure 2
Profiles of expression of brain derived neurotrophic factor in serum and cerebrospinal fluid. (A) Immunoblot analysis of brain derived neurotrophic factor (BDNF) isoforms in serum disclosed the presence of the pro-form (32 kD), truncated-form (28 kD) and mature form (14 kD) while plasma disclosed mostly the presence of the BDNF pro-form and truncated-form. The truncated-BDNF form was the only isoform seen in cerebrospinal fluid (CSF). (B)The BDNF truncated-form was significantly decreased after treatment with minocycline (P = 0.028) a trend that was observed in six of ten patients (red lines). (C) Although the mature form appeared to be decreased after treatment the difference pre- and post-treatment did not reach significance (P = 0.38). (D) BDNF in CSF measured by multiplexed array technique, which detects mostly the truncated-BNDF isoform, was significantly lowered after treatment (P = 0.042).
Figure 3
Figure 3
The levels of hepatic growth factor (HGF) in cerebrospinal fluid (CSF) were significantly increased following treatment with minocycline. (P = 0.028).
Figure 4
Figure 4
A post-treatment lowering effect on MMP-7 was observed in seven of ten patients (red lines), but despite this trend it did not reach statistical significance. (P = 0.059).

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