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4/8/2016 Copyright by Dr. TC Theoharides 1

Theoharis C. Theoharides, MS, MPhil, PhD, MD, FAAAAI

Professor of Pharmacology, Internal Medicine and Psychiatry

Director, Molecular Immunopharmacology and Drug DiscoveryTufts University School Medicine, Boston, MA, USA

drtheoharides@gmail.comwww.mastcellmaster.comwww.autismfreebrain.org

www.brain-gate.org

14/8/2016 Copyright by Dr. TC Theoharides

An Autism Subgroup With Brain Inflammation

Is Treatable

Disclosures

• Scientific Director and shareholder of Algonot, LLC

• Medical Advisory Board, The Mastocytosis Society

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Autism Spectrum Disorders

Rett Syndrome

Autistic Disorder

Childhood Disintegrative

Disorder

PDD-NOS* Asperger’s SyndromePDD‐NOS, pervasive 

developmental  disorder not otherwise specified.

• ADD/ADHD• Allergies/Food intolerance• Gastrointestinal problems• Mitochondrial dysfunction• PANDAS (Pediatric Autoimmune Neuropsychiatric

Disorders Associated With Streptococcal Infections)• PTEN mutations (large skull, headaches)• Seizures

Subgroups

K. Weintraub, Nature 479, Nov. 3 2011, 22-24.

Deficits in communication and sociability together with repetitive movements

Now 1 in 45 children

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Presenting Behavior Suggestive of Autism

• No response when calling name• Isolation• No group play• Loss of vocabulary• Rigid routines• Lack of imaginative play• Inability to follow directions• Anxiety• Intolerance to stress• Repetitive non-purposeful

movements• Hyperactivity• Tantrums• Tip-toeing• Hand flapping• Intolerance to sensory overload• Strange food habits

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Gestational and Perinatal Factors Increasing Autism Risk

Gestational

• Allergies/Asthma• Autoimmune diseases• Diabetes• Infection• Low Vitamin D• Pre-eclampsia• Psoriasis• Psychotropic drug use • Stress

Perinatal• Excessive oxytocin to induce labor

• High fever

• Incubation/maternal separation stress

• Infection

• Low birth weight

• Perinatal anoxia

• Preterm birth

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Diencephalon(coordinates sensory input and emotions)

Mast Cell Activation

Dysfunctional social behavior, communication and stereotypic

movements

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There was significant association with food intolerance (P = 0.001).

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Croen et al. Arch Pediatr Adolesc Med 2005;159:151-7.

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Autism Spectrum Disorders and MastocytosisTheoharides TC. Int J Immunopathol Pharmacol.

2009 Oct-Dec;22(4):859-65.The risk of autism was 10 times higher than in the general population.

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Is There Evidence of Inflammation in the Brain?

• Activation and proliferation of microglia, the brain immune cells, causes local inflammation and “choking” of neuronal connectivity

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IL-6, TNFMicroglia

Neuron

Mast cell

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How Can Stress Contribute to Inflammation of the Brain?

• Disrupts the gut-blood barrier

• Opens up the blood-brain barrier

• Stimulates mast cells

• Triggers microglia cells

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Prenatal Stress Has Been Associated With High Risk of Developing Autism

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tv 99Tc-G

30 minutes restraint stress

C57BL +/+ normalW/Wv mast cell-deficient

mouse

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0

50

100

150

200

*

+/+ W/Wv%

ch

ang

e fr

om

co

ntr

ol

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Intradermal CRH Injection Induces Skin Vascular Permeability, but not in W/Wv Mast Cell Deficient Mice

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Culture of human umbilical cord blood-derived mast cells (hCBMCs)

Culture with SCF (100 ng/ml) and IL-6 (50 ng/ml) for 8-16 wk yields 100% tryptase-positive mast cells

CD34+ cells

mast cellsKempuraj D., Saito H. et al. Blood 93: 3338-3346, 1999.

RBCs

MNCs

AC133 mAb

MACs

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Immunocytochemistry of CRH-R in human mast cells

F G

B

-

R1 no IL-4

DCA E

E F

B

R1 + IL-4

A B

R2 no IL-4 R2 + IL-4

DB

No R1 primary DAPI

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Effect of Mast Cells on the HPA Axis

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The Peptide Neurotensin Can Trigger Inflammation of the Brain

• Is increased in the blood of children with ASD

• Is found in the brain and gut

• Stimulates mast cells and microglia

• Has synergistic action with CRH

• Opens up the blood-brain barrier

• Is neurotoxic

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What Triggers Inflammation in the Brain?

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01020304050

control 1nM 10nM 100nM 1uMCRH

VE

GF

A) B)

C) D)

Figure 3 Microglia Express NTR3/Sortilin, which is Elevated in the Serum of Children with ASD

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Neurotensin Stimulates Human Microglia IL1 Gene Expression and Release

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Treatment

Beaudet. Science; 338:342-3, 2012

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Wong AY et al., Autism Res. 2014 Oct;7(5):543-54

Most Countries Use Psychotropic Drugs in Children with ASDs

Luteolin (3′,4′,5,7-tetrahydroxyflavone)

Methoxyluteolin (3′,4′,5,7-tetramethoxyluteolin)

Properties of Luteolin Useful in Autism

• Reduces oxidative stress• Inhibits inflammation• Inhibits mast cell activation• Inhibits microglia activation• Inhibits neurotoxicity• Is weak metal chelator• Increases memory• Prevents autism-like behavior in mice

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0

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20

30

40

50

60

70

80

90

100

Histamine Tryptase IL-6 IL-8 TNF-alpha

Per

cen

t in

hib

itio

n

*

** *

*

Inhibition of inflammatory molecules by quercetin from human mast cells

Effect of luteolin on SP-induced human mast cell secretion

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Jang et al. Proc Natl Acad Sci USA, 2008;105:7534-9

Luteolin significantly inhibitedIL-6-induced behavioral deficits in social interactions in mice

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Methoxyluteolin Inhibits Inflammatory Molecule Release from Human Microglia

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Methoxyluteolin Inhibits IL-33/SP-Induced TNF Secretion and NFB Activation in Mast Cells

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Mast Cell Triggers

PI3K

AKT

mTORC1

PTENMast cell number

mast cell activation

Mast cell number

Rapamycin

Luteolin

Calcium influx via receptor

Methoxyuteolin inhibits mTOR activation

1-Control 2-Rapamycin 3-Methoxyluteolinteolin

MAST CELL SECRETION

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A novel, hypoallergenic, anti-oxidant, flavonoid lotion for dry, irritated or sensitive skin

Hydrates-Nourishes–Soothes-Rejuvenates-RepairsLet Nature Calm Your SkinTM

Net weight 2 oz (56.7 gm)

Ingredients: Water, organic aloe barbadensis leaf juice, olea europea (olive) fruit oil, glycerin, organic helianthus annuus (sunflower) seed oil, steareth-21, stereath-2, cetyl

alcohol, stearyl alcohol, glyceryl stearate, lanolin, tetramethoxyflavone, sodium benzoate, quaternium-15, disodium EDTA, magnesium chloride, honey, sodium chloride, potassium

chloride, tetrahexyldecyl ascorbate, origanum vulgare oil, disodium phosphate

Made for Algonot, LLC, 5111 Ocean Bvd-Suite J Sarasota, FL 34242, USA-Toll Free 1-800-254-6668

www.gentlederm.com

GentleDermTM

A B C D

Photos of lesional skin areas. Areas were photographed before (upper panels) and after (lower panels) using GentleDerm® twice daily for two weeks. (A,B) eczema: (A) face of a 51 year-old female patient with

mast cell activation syndrome and (B) abdomen of 43 year-old female patient with mast cell activation syndrome;(C,D) psoriasis: (C) leg of a 47 year-old male patient with indolent systemic mastocytosis and (D) hand of

39 year-old female patient with mast cell activation syndrome.

Mast Cell

Microglia

Inflammatory

mediators

Neurons

Neuronal damage and impaired connectivity

H

No trigger Triggers +Atopy + CRH +Neurotensin + mTOR activation

Triggers +Atopy

Triggers

INSCREASED RISK OR AUTISMTriggers +Atopy + CRH+Neurotensin

Lut & Methlut

Treatment Approaches• Reduce anxiety/pressure

• Limit sensory overload

• Start speech, music and work support

• Correct vitamin deficiencies

• Supplement methyl donors (glutathione, SAMe, NAC)

• Eliminate food intolerances

• Reduce allergies

• Add anti-oxidants (Omega-3, sulforaphane)

• Address inflammation (luteolin, methoxyluteolin)

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Research Funding:

• DK62861 • AR47652 • NS071361 • NS55681• AR60951• NS66205

• Autism Research Collaborative

• National Autism Association

• Safe Minds• Autism Research

Institute• Jane B Johnson Fnd.

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drtheoharides@gmail.comwww.mastcellmaster.comwww.autismfreebrain.org

www.brain-gate.org