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. 2025 Aug 26;26(17):8293.
doi: 10.3390/ijms26178293.

Transgenerational Effects and Heritability of Folate Receptor Alpha Autoantibodies in Autism Spectrum Disorder

Richard E Frye  1 , Ira L Cohen  2 , Jeffrey M Sequeira  3 , Zoe Hill  1 , Alina Espinoza  1 , W Ted Brown  4 , Clifford Mevs  5 , Elaine Marchi  4 , Michael Flory  6 , Edmund C Jenkins  4 , Milen T Velinov  7 , Edward V Quadros  3
Affiliations

Transgenerational Effects and Heritability of Folate Receptor Alpha Autoantibodies in Autism Spectrum Disorder

Richard E Frye et al. Int J Mol Sci. .

Abstract

Autism Spectrum Disorder (ASD) affects an estimated prevalence of 1 in 31 children but the cause in most cases is unknown. Human and animal studies have linked ASD to Folate Receptor Alpha Autoantibodies (FRAAs). Our previous studies demonstrated that FRAAs are more common, on average, in families with children with ASD. This study reanalyzed data from a previous study which included 82 children diagnosed with ASD, 53 unaffected siblings, 70 mothers, 65 fathers, and 52 typically developing controls who did not have a history of ASD in their family. This study investigates the association of FRAA titers with ASD risk factors and explores the relationship of FRAA titers across generations. Several known risk factors for ASD, including multiplex ASD families, multiple birth pregnancies, and increased maternal and paternal ages at birth, were related to offspring FRAA titers. Multiplex ASD families demonstrated higher FRAA titers. Significant correlation were found between maternal and offspring blocking FRAA titers. FRAA titers increased across generations, although the increase in blocking FRAA titers was only seen in multiplex families. The proband with ASD showed higher blocking but not higher binding, FRAA titers compared to their non-affected siblings. Paternal FRAA titers are associated with several measures of offspring behavior and cognitive development. This research highlights the potential transgenerational transmission of FRAAs and their role in ASD. This supports the notion that heritable non-genetic factors may be important in the etiology of ASD and that FRAAs may demonstrate anticipation (worsening across generations), especially in multiplex families. FRAAs may provide one example of the possibility that susceptibility to autoimmune processes may contribute to disrupted brain development and function in ASD.

Keywords: anticipation; autism spectrum disorder; cerebral folate deficiency; folate receptor alpha; folate receptor alpha autoantibodies; heritability.

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Conflict of interest statement

E.V.Q. and J.M.S. are inventors on a US patent for the detection of FRAAs issued to the Research Foundation of the State University of New York, NY, USA. I.L.C. was involved in the development of the PDDBI and receives royalties from the use of this instrument. The remainder of the authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Cellular folate metabolism. The folate cycle is central to other critical cellular biochemical systems including methylation, glutathione production, and neurotransmitter production. The key enzymes for biochemical reactions are provided in the rectangles, with the enzymes having polymorphism overrepresented in ASD colored in purple. 5-MTHF/CH3-THF: 5-methyltetrahydrofolate; B12: Vit B12 (cobalamin); BH4: tetrahydrobiopterin; BHMT: Betaine-homocysteine methyltransferase; CBS: cystathionine beta-synthase; CH2-THF: 5,10-methylenetetrahydrofolate; COMT: Catechol-O-methyltransferase; CTH: cystathionine gamma-lyase; DHFR: dihydrofolate reductase; DNA: deoxyribonucleic acid; DNMT1: DNA methyltransferase 1; DNMT3A: DNA methyltransferase 3A; GCH1: GS: glutathione synthetase; GTP cyclohydrolase I; MAT: methionine adenosyltransferase; MBD4: Methyl-CpG-binding domain protein 4; Me: methyl group; MTR: Methionine synthase; MTHFD1: Methylenetetrahydrofolate dehydrogenase 1; MTHFD2: Methylenetetrahydrofolate dehydrogenase 2; MTHFR: methylenetetrahydrofolate reductase; MTRR: methionine synthase reductase; RNA: ribonucleic acid; SAHH: S-adenosylhomocysteine hydrolase; SHMT: Serine hydroxymethyltransferase; TCN2: Transcobalamin II; THF: tetrahydrofolate. Created in BioRender.
Figure 2
Figure 2
The transport of folate across the placenta and into the nervous system and the role of folate receptor alpha (FOLR1) autoantibodies (FRAAs) in disrupting this transport. F: folate, PCFC: protein coupled folate carrier, RFC: reduced folate carrier. Created in BioRender.
Figure 3
Figure 3
The relationship between age and FRAA titers. (A) Both maternal (blue line) and paternal (red line) ages were related to blocking FRAA titers such that higher levels of blocking FRAA titers in offspring were associated with a higher parental age. (B) Binding FRAA titers decreased with older offspring ages.
Figure 4
Figure 4
The relationship between offspring and maternal blocking FRAA titers. The blue circles represent multiplex families while the orange squares represent simplex families. Oral highlights the cases in which both offspring and maternal blocking FRAA titers were positive. These cases are disproportionally from multiplex families.
Figure 5
Figure 5
Relationship of FRAA titers between parents and offspring. The change in blocking titers across generations was significantly higher in (A) multiplex as compared to (B) simplex families. For both multiplex and simplex families, offspring blocking titers were significantly higher than parental blocking titers. (B) The titers for the offspring, mothers, and fathers were similar in simplex families. (C) Binding titers were not significantly difference across family members.
Figure 6
Figure 6
Comparison of (A) blocking and (B) binding FRAA titers in proband families and typically developing controls.
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