Single Cell ADNP Predictive of Human Muscle Disorders: Mouse Knockdown Results in Muscle Wasting

Abstract

Activity-dependent neuroprotective protein (ADNP) mutations are linked with cognitive dysfunctions characterizing the autistic-like ADNP syndrome patients, who also suffer from delayed motor maturation. We thus hypothesized that ADNP is deregulated in versatile myopathies and that local ADNP muscle deficiency results in myopathy, treatable by the ADNP fragment NAP. Here, single-cell transcriptomics identified ADNP as a major constituent of the developing human muscle. ADNP transcript concentrations further predicted multiple human muscle diseases, with concentrations negatively correlated with the ADNP target interacting protein, microtubule end protein 1 (EB1). Reverting back to modeling at the single-cell level of the male mouse transcriptome, Adnp mRNA concentrations age-dependently correlated with motor disease as well as with sexual maturation gene transcripts, while Adnp expressing limb muscle cells significantly decreased with aging. Mouse Adnp heterozygous deficiency exhibited muscle microtubule reduction and myosin light chain (Myl2) deregulation coupled with motor dysfunction. CRISPR knockdown of adult gastrocnemius muscle Adnp in a Cas9 mouse resulted in treadmill (male) and gait (female) dysfunctions that were specifically ameliorated by treatment with the ADNP snippet, microtubule interacting, Myl2—regulating, NAP (CP201). Taken together, our studies provide new hope for personalized diagnosis/therapeutics in versatile myopathies.

A novel davunetide (NAPVSIPQQ to NAPVSIPQE) point mutation in activity‐dependent neuroprotective protein (ADNP) causes a mild developmental syndrome

“…This is translated into altered cellular dynamics and reflected in developmental effects on transient eating disorders (vomiting and low weight gain, digestions problems), and muscle (potentially neuromuscular junction) deficits, as well as mild behavioural deficits. These findings are in agreement with animal studies on ADNP deficiencies (Hacohen-Kleiman et al, 2018;Kapitansky, Karmon et al, 2020; and extended to ADNP truncating mutations (Figure 2), (Karmon et al, 2022).…”

“…These are coupled to NAP enhancement of ADNP interaction with microtubule‐associated protein 1 light chain 3 (LC3) (Merenlender‐Wagner et al, 2015) constituting the autophagosome membrane, indicative of control of fundamental cellular processes. These interactions are further associated with ADNP/NAP control of gene expression central to brain/body functions (Amram et al, 2016; Hacohen‐Kleiman et al, 2018, 2019; Kapitansky, Karmon et al, 2020; Kapitansky, Sragovich et al, 2020; Karmon et al, 2022; Sragovich et al, 2019).…”

“…In the ADNP child (B) the mutation was mostly associated muscle‐related problems as well as some social problems, related to interactions with peers. Thus, a new face of the ADNP syndrome is revealed, with ADNP strongly implicated with muscle function (Kapitansky, Karmon et al, 2020; Kapitansky, Sragovich et al, 2020).…”

Chromatin remodeler Activity-Dependent Neuroprotective Protein (ADNP) contributes to syndromic autism

“…The demonstrated sexual dichotomy of Adnp haploinsufficient mice [ 23 , 98 , 105 , 122 ] is further enhanced in the Tyr-mouse. Developmental delays and gait dysfunctions were increased in Tyr females compared to Adnp ± mice, with adult muscle knockdown of Adnp resulting in robust female gait defects, corrected by NAP [ 134 ], coupled with sex-muscle-specific differentially expressed genes, also partly corrected by NAP [ 135 ]. The mouse models suggest peripheral biomarkers, like FOXO3, and with FOXO3 regulating the microbiome, the ADNP/NAP peripheral biomarkers include a microbiota signature [ 136 ].…”

Tau, ADNP, and sex

“…These findings are further translated into sex-dependent environmental sensation (auditory and visual responses (Hacohen-Kleiman et al, 2019;Karmon et al, 2022), motor function (Kapitansky, Karmon, et al, 2020;Karmon et al, 2022) and behavior (Malishkevich et al, 2015;Sragovich et al, 2019). Just touching the tip of the iceberg, in terms of gene expression, ADNP/NAP regulate β3 tubulin expression (Oz et al, 2012), associated with neuronal maturation (Gozes & Littauer, 1978;Gozes & Sweadner, 1981), with further sex-dependent regulation of tubulin isotypes (Amram et al, 2016) linked with brain development (Gozes & Littauer, 1978).…”