Addressing gastrointestinal distress, neuroinflammation and immune modulation in ASD patients via supplementation with mushroom nutrition

Autism Spectrum Disorder (ASD) is a disorder still very poorly understood first recognized in early childhood in the form of a multi organ system disability caused by impaired neurogenesis and apoptosis, impaired synaptogenesis and synaptic pruning or imbalanced excitatory-inhibition system (1). 

Inflammation has been recognised as the pathogenesis of autism, but a holistic approach is required. The aetiology is largely unknown and there is no clinical treatment. The gut microbiota may affect symptom manifestation which may benefit from a balanced diet, re-establishment of intestinal permeability, improvement of gut microbiota, raised immunity, supply of antioxidants and detoxification speed (1). 

The gastrointestinal (GI) comorbidities, including acute and chronic constipation and diarrhoea as well as persistent neuroinflammation are well documented. (2-7)

The following outlines the concept of the overlap syndrome of GI disorders and ASD (8).

Figure 1.  The concept of the overlap syndrome of GI disorders and ASD.

ASD patients also have low presence of Natural Killer cell activity (NK cell) (9). This immunological dysfunction may explain the higher cancer mortality rates in ASD patients (10).  

Clinical research over the past seven years have reinforced the possibility that mushroom nutrition is a possible disease modifying therapy for ASD patients by addressing the comorbidities involving gastrointestinal distress, neuroinflammation and reduced immune status. The research has focused on supplementation with two forms of mushroom biomass, Coriolus versicolor and Hericium erinaceus.

Addressing ASD Comorbidities

A. Gastrointestinal Distress

Coriolus versicolor (‘Turkey Tail’) is a mushroom known for its immunomodulatory properties (11) but has also displayed prebiotic-like activity (12-13), which has been attributed to its specific β-glucans (a naturally occurring glucose polysaccharide) (12).

In 2018, researchers at the Catholic University of Porto, Portugal evaluated the prebiotic potential of Coriolus versicolor biomass through in vitro faecal fermentations, using fresh faecal samples from five healthy donors, and performed subsequent analysis on changes in the bacterial population and the production of Short Chain Fatty Acids (SCFA).

The researchers found:

1. Coriolus versicolor biomass induced changes in the gut microbiota. There was enhanced growth of potentially beneficial bacterial (Bifidobacterium) populations vs negative controls. The effect was similar, if less intense, to that observed with a positive control containing a well-known prebiotic, fructo-oligosaccharides (FOS) in an in vitro system (11).
2. The pH values decreased throughout the 48 h fermentation, indicating that Coriolus versicolor was fermented and resulted in the production of SCFA, at lower levels than those of the positive control (11).
3. The fermentation led to SFCA production and glucose consumption (11). 

 In 2022, a study suggested that the mushroom species Coriolus versicolor (CV) may have therapeutic potential for the prevention and treatment of inflammatory bowel diseases. (14).

Using a mouse colitis model, the study found that CV biomass reduced histological signs of damage, attenuated the rise in multiple markers of inflammation, and boosted anti-inflammatory responses (14). 

Researchers from collaborating universities in Italy and the US induced colon damage in mice by intrarectally administering dinitrobenzene sulphonic acid (DNBS). Four groups of 10 mice were used: 

a) Sham (intrarectal administration of 100 µL 50% ethanol); 

b) Sham + Coriolus versicolor (200 mg/kg dissolved in saline, orally 1 h after sham procedure every 24 h); 

c) DNBS-treated (4 mg in 100 µL of 50% ethanol per mouse); and

d) DNBS-treated + CV (200 mg/kg dissolved in saline, orally 1 h after DNBS procedure and every 24 h). 

Four days after the procedures, the mice were sacrificed. 

Effects on physical appearance of the colon, weight loss and colon physiology (14)

• Physical appearance
  The colons of the sham groups appeared normal. Those of the untreated colitis model group were flaccid, with liquid stools and, in some cases, were ulcerated with mucosal congestion. These changes were significantly less in the CV-treated colitis model group.
• Body weight
  The significant loss of body weight (caused by diarrhea) in the colitis model group was significantly reduced in the CV-treated colitis model group. 

• Physiology
  Histological analysis of sections of colon tissue revealed that DNBS had induced infiltration of inflammatory cells (mostly neutrophils – also see later), necrosis and oedema; although present in the CV-treated colitis model group, these pathological changes were significantly less marked.

Effects on inflammatory processes (14)

The effects of CV in preventing inflammation of the colon were investigated by assessing multiple markers of increased inflammation. 

• TLR4/NFĸB inflammatory pathway

Immunohistochemical analysis indicated that expression of the receptor TLR4 increased in the untreated colitis model mice, as did Myd88 expression, degradation of IĸB-α and nuclear translocation of NF-ĸB – all components involved in the TLR4/NFĸB inflammatory pathway. Together, these changes indicated upregulation of the inflammatory process. These pro-inflammatory effects were significantly limited in the CV-treated colitis model mice.

• Infiltration of neutrophils and lipid peroxidation

Neutrophil infiltration into the colon was assessed by measuring myeloperoxidase levels; the extent of lipid peroxidation in the colon was determined by activity of malondialdehyde. The large increases in these parameters (both of which suggest a pro-inflammatory status) in untreated colitis model mice were significantly attenuated in the CV-treated colitis model mice. 

• Activation of T cells and adhesion molecules 

The number of CD4+ and CD8+ (T cells that are important mediators of inflammation), and the expression of the adhesion molecules P-selectin (a transmembrane protein that has an essential role in inflammatory response to injury) and ICAM1 (a cell surface glycoprotein involved in recruiting active leucocytes to sites of inflammation) was assessed using immunohistochemical tissue staining. All parameters were significantly increased in the untreated colitis model mice, while their rise in CV-treated colitis model mice was significantly limited.

• Proinflammatory mediators
  ELISA testing showed that proinflammatory mediators – CCL2, PGE2, IL-1β and TNF-α – were raised in the untreated colitis model mice, but significantly less-so in the CV-treated colitis model mice.

• Nitrosative stress and PARP hyperactivation
  The levels of nitrosative stress and extent of PARP activity in colon samples – both indicative of potential for cell damage – were assessed immunohistochemically. There was a high level of positive staining of samples taken from colitis model mice (indicating a high potential for cell damage), with significantly less staining (and thus lower potential for damage) in samples from the CV-treated colitis model mice.

Enhancement of anti-inflammatory pathways (14)

• Reactive oxygen species are important potentiators of inflammation; and Nrf2 is a protein responsible for controlling the expression of genes encoding enzymes that detoxify and eliminate reactive oxygen species (known as HO-1 enzymes). Western blot analysis of colon samples revealed that colitis model mice that had consumed CV had a much higher expression of Nrf2 and HO-1 than mice that had not consumed CV (both sham-operated and colitis model); this indicated that CV-treated mice were better able to ameliorate the effects of oxidative stress.

These results suggest that Coriolus versicolor supplementation hold great promise as a useful way to manage inflammatory bowel diseases and to support the vital Gut-Brain Axis by acting as a prebiotic and reducing gastrointestinal inflammation.

Host-Microbial Gut Interactions and Mushroom Nutrition. Victoria Bell, Jorge Ferrão, Eusébio Chaquisse, Tito Fernandes. Journal of Food and Nutrition Research. 2018;6(9):576-583. doi:10.12691/jfnr-6-9-6.

B. Reducing Inflammation and Neuroinflammation

Increasing evidence indicates that aspirin-triggered Lipoxin A4 (LXA4) (15 μg/kg) s c, twice a day, reduced both NF-kB activation and levels of proinflammatory cytokines and chemokines, as well as increased levels of anti-inflammatory IL-10 and transforming growth factor B (beta). Basically, LXA4 reduces inflammation. (15).

Working with Sprague-‐Dawley rats, researchers at the University of Catania, Italy determined that at when the rats were supplemented separately with Corious versicolor (2015) and Hericium erinaceus (2016), -there was a significant increase in Lipoxin A4 within 30 days and 90 days respectively (16, 17). 

At the end of experimental period animals were sacrificed and the activity of LXA4 was determined in serum, lymphocytes and in different brain regions (cortex, striatum, substantia nigra, hippocampus and cerebellum) and compared with LXA4 of untreated animals, as control (16,17). 

The researchers also focused on the impact of Coriolus versicolor and Hericium erinaceus (“Lion´s Mane) on the differences in the up-regulation of Heme Oxygenase-1 (HO-1), Heat Shock Protein 70 (Hsp 70) and Thioredoxin (TrX).

Figure 3. LXA4 distribution in the brain of Sprague-Dawley rats after supplementation with Coriolus versicolor (30 days)  and Hericium erinaceus (90 days) 

Figure 4. LXA4 distribution in the rest of body of Sprague-Dawley rats after supplementation with Coriolus versicolor (30 days)  and Hericium erinaceus (90 days)

Figure 5. HO-1, HSP70 and TrX distribution in the brain of Sprague-Dawley rats after supplementation with Coriolus versicolor (30 days)  and Hericium erinaceus (90 days)

In a 2018-2019 follow-up study, the same University of Catania researchers, enrolled 40 patients with Meniere’s disease (MD) of which 22 were supplemented with Coriolus versicolor biomass (3 g/day-6 tablets per day-3 tablets in morning and 3 tablets in evening) over two months and 18 participants were part of the control group (18).

To date, the cause of MD remains largely unknown, although increasing evidence suggests that, as an oxidant disorder, oxidative stress, immunomodulation and neuroinflammation may be central to its pathogenesis (18).  

This study noted not only an increase in Lipoxin A4 levels but a reversal of both oxidative and cellular stress response in these patients which was reflected beneficial changes in their symptom scores and improved biomarkers after 60 days (18). 

Figure 6: Levels of LXA4 in the plasma (a), lymphocytes (b) and urine (c) of treated (22) and untreated (18) MD patients (estimated by HPLC + mass spectrometry). *p < 0.05 vs untreated MD

C. Improving Immune System

As previously stated, ASD patients also have low presence of Natural Killer cell activity (NK cell) (9). This immunological dysfunction may explain the higher cancer mortality rates in ASD patients (10).  

It has been noted in a chronic fatigue syndrome study by Dr.Jean Monro that Coriolus versicolor can increase Natural Killer cell activity by 35% within 8 weeks of supplementation (19). Supplementation was 6 tablets (3 g) per day for two weeks followed by 3 tablets (1.5 g) per day for six weeks. 

In addition, Coriolus versicolor has been used in LSIL HPV clinical trials.  In a one year clinical trial, Coriolus versicolor supplementation (3g/day-6 tablets) was found to promote cervical lesion regression rate of 72% vs 47% in a control group, while the HPV virus was undetectable in 90% of the patients versus 8% in the control group over the same time period (20).

In summary, the individual profiles of Coriolus versicolor and Hericium erinaceus (Hericor-MRL) biomass demonstrate the ability to modulate the microbiome, reduce both inflammation and neuroinflammation, while providing an immune modulation protection. This set of attributes has potential applications in addressing the comorbidities in ASD patients. 

Mushroom Nutrition Supplementation to Modulate the Immune System in ASD Patients

References

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(11) Cruz A, Pimentell, L, Rodríguez-Alcalá LM, Fernandes T, Pintado M. Health benefits of edible mushrooms focused on Coriolus Versicolor: a review. Journal of Food and Nutrition Research 2016;4(12):773781. doi: 10.12691/jfnr4122.

(12) Yu Z-T, Liu B, Mukherjee P, Newburg DS. Trametes versicolor extract modifies human fecal microbiota composition in vitro. Plant Foods for Human Nutrition 2103;68(2):107-112. doi: 10.1007/s11130-013-0342-4. 

(13)  Wang B, Yao M, Lv L, Ling Z, Li L. The human microbiota in health and disease. Engineering 2017;3:71-82.  https://doi.org/10.1016/J.ENG.2017.01.008.

(14) Impellizzeri D, Fusco R, Genovese T, Cordaro M, D’Amico R, Trovato Salinaro A, Ontario ML, Modafferi S, Cuzzocrea S, Di Paola R, Calabrese V, Siracusa R. Coriolus Versicolor Downregulates TLR4/NF-ĸB Signaling Cascade in Dinitrobenzenesulfonic Acid-Treated Mice: A Possible Mechanism for the Anti-Colitis Effect. Antioxidants 2022;11:406. https://doi.org/10.3390/antiox11020406

(15) Medeiros R, Kitazawa M, Passos GF, Baglietto-Vargas D, Cheng D, Cribbs OH, LaFerla FM. Aspirin-triggered lipoxin A4 stimulates alternative activation of microglia and reduces Alzheimer disease-like pathology in mice. Am J Pathol. 182(5);1740-9. May 2013.

(16) Trovato A, Siracusa R, Di Paola R, Scuto M,Fronte V, Koverech C, Luca M, Serra A, Toscano M.A., Petralia A, Cuzzocrea S, Calabrese V. Redox modulation of cellular stress response and lipoxin A4 expression by Coriolus versicolor in rat brain: Relevance to Alzheimer´s disease pathogenesis. Neurotoxicology. 53:350-8. doi: 10.1016/j.neuro.2015.09.012. 2016.

(17) Trovato A, Siracusa R, Di Paola R, Scuto M, Ontario ML, Bua O, Di Mauro P, Toscano MA, Petralia CC, Maiolino L, Serra A, Cuzzocrea S, Calabrese V. Redox modulation of cellular stress response and lipoxin A4 expression by Hericium erinaceus in rat brain: relevance to Alzheimer’s disease pathogenesis. Immun Ageing. 13:23. doi: 10.1186/s12979-016-0078-8. Jul 9 2016.

(18) 17. Scuto, M.; Di Mauro, P.; Ontario, M.L.; Amato, C.; Modafferi, S.; Ciavardelli, D.; Trovato Salinaro, A.; Maiolino, L.; Calabrese, V. (2020). Nutritional Mushroom Treatment in Meniere’s Disease with Coriolus versicolor: A Rationale for Therapeutic Intervention in Neuroinflammation and Antineurodegeneration. Int. J. Mol. Sci. 2020, 21, 284. doi: 10.3390/ijms21010284

(19) Monro J.A, Coriolus  J Integrative Medicine 2004;8:101-108.

(20) Couto, J.S,  Salgueiro, L Evaluation of the Efficacy of Coriolus versicolor in the Treatment of HPV Lesions (LSIL). Poster presentation at the 14^th World Congress of Cervical Pathology and Colposcopy-IFPC. July 7^th, 2011 -Rio de Janeiro, Brazil.