Expansion of the phenotypic spectrum of de novo missense variants in kinesin family member 1A (KIF1A).

Defects in the motor domain of kinesin family member 1A (KIF1A), a neuron-specific ATP-dependent anterograde axonal transporter of synaptic cargo, are well-recognized to cause a spectrum of neurological conditions, commonly known as KIF1A-associated neurological disorders (KAND). Here, we report one mutation-negative female with classic Rett syndrome (RTT) harboring a de novo heterozygous novel variant [NP_001230937.1:p.(Asp248Glu)] in the highly conserved motor domain of KIF1A. In addition, three individuals with severe neurodevelopmental disorder along with clinical features overlapping with KAND are also reported carrying de novo heterozygous novel [NP_001230937.1:p.(Cys92Arg) and p.(Pro305Leu)] or previously reported [NP_001230937.1:p.(Thr99Met)] variants in KIF1A. In silico tools predicted these variants to be likely pathogenic, and 3D molecular modeling predicted defective ATP hydrolysis and/or microtubule binding. Using the neurite tip accumulation assay, we demonstrated that all novel KIF1A variants significantly reduced the ability of the motor domain of KIF1A to accumulate along the neurite lengths of differentiated SH-SY5Y cells. In vitro microtubule gliding assays showed significantly reduced velocities for the variant p.(Asp248Glu) and reduced microtubule binding for the p.(Cys92Arg) and p.(Pro305Leu) variants, suggesting a decreased ability of KIF1A to move along microtubules. Thus, this study further expanded the phenotypic characteristics of KAND individuals with pathogenic variants in the KIF1A motor domain to include clinical features commonly seen in RTT individuals.

Age and sex influences on airway hyperresponsiveness.

OBJECTIVE: The precise relationship between age and gender and their influence on airway reactivity has not been clearly defined. Previous studies of age and gender influences on airway reactivity have been confounded by environmental influences such as cigarette smoking. The objective of this study was to examine the effect of age and gender on airway reactivity in C57BL/6 mice housed under controlled conditions, independent of confounding environmental factors. METHODS: Mice were separated into four experimental groups based on age and gender: males at 6 and 12 weeks of age and females at 6 and 12 weeks of age. Airway reactivity to inhaled methacholine was examined in each group. RESULTS: Significant differences in methacholine-induced airway reactivity were observed between the sexes at both age groups. At 6 weeks of age, the males demonstrated a significantly increased airway reactivity to methacholine as compared to females (p <.001). Paradoxically, at 12 weeks of age, the males demonstrated a significantly lower response to methacholine as compared to females (p <.001). Hence the relationship between age and airway reactivity is markedly different in males as compared to females. Examination of intragender differences revealed that in female mice, airway reactivity increased significantly with age (p <.001). In contrast, males demonstrated a significant reduction in airway reactivity with age (p <.001). CONCLUSION: These findings demonstrate important differences in airway reactivity related to age and gender that are observed independent of any environmental influences. Furthermore, these findings highlight the importance of careful age and sex matching in studies of airway reactivity.

Attenuated methacholine airway response following repeat testing in a murine model of allergic airways disease.

A progressive attenuation of airway reactivity to methacholine is observed in normal individuals with successive bronchial provocation testing. The absence of this attenuation in asthma is thought to be due airway inflammation. The authors investigated this phenomenon in a mouse model of allergic airways disease. Repeated measurements of airway response were carried out in mice sensitized/challenged with ovalbumin or saline, and in untreated mice. Saline-treated and untreated mice showed reduced airway reactivity following repeated testing. This was also observed in ovalbumin-treated mice in the second and third tests compared to the previous test (P < .05). This attenuation was not associated with airway inflammation, which remained high in the ovalbumin group. The results suggest that attenuation of airway reactivity with repeated methacholine challenge is due to factors other than airway inflammation.

Relaxin plays an important role in the regulation of airway structure and function.

Relaxin is a reproductive hormone with pleiotropic actions. In addition to airway fibrosis, relaxin deficiency results in airway structural changes (epithelial thickening) and increased lung recoil, suggesting that relaxin may impact other aspects of airway/lung structure and function beyond its ability to regulate collagen turnover. Furthermore, these structural changes associated with relaxin deficiency show marked similarity to the structural changes seen in asthma. The current study investigated the broader role of relaxin in regulating airway structure and function and examined the relationship between airway inflammation, structural changes, and airway hyperresponsiveness (AHR) using an ovalbumin (OVA)-induced model of allergic airways disease (AAD). The model of AAD was applied to 12-month-old relaxin-deficient (Rln(-/-)) mice with established airway fibrosis and age-matched wild-type (Rln(+/+)) controls. OVA-treated Rln(+/+) mice (induced inflammation) developed increased epithelial thickening (P < 0.05) and AHR (P < 0.05) but not airway fibrosis, compared with saline-treated Rln(+/+) controls. Saline-treated Rln(-/-) mice had significantly increased lung collagen deposition (existing fibrosis) and epithelial thickening and remarkably were found to have increased AHR that was equivalent to that in OVA-treated Rln(+/+) mice (all P < 0.05 vs. saline-treated Rln(+/+) controls). OVA-treated Rln(-/-) mice (existing fibrosis and induced inflammation) had increased airway/lung fibrosis (P < 0.05) but equivalent airway inflammation and AHR compared with OVA-treated Rln(+/+) animals. These findings demonstrate for the first time a role for relaxin in the regulation of airway responses using Rln(-/-) mice and suggest that airway fibrosis and/or epithelial thickening can result in increased AHR equivalent to that induced by airway inflammation in AAD.

Relaxin deficiency in mice is associated with an age-related progression of pulmonary fibrosis.

Relaxin (RLX) is a peptide hormone with known antifibrotic properties. However, its significance in the lung and its role as a therapeutic agent against diseases characterized by pulmonary fibrosis are yet to be established. In this study, we examined age-related structural and functional changes in the lung of relaxin-deficient mice. Lung tissues of male and female RLX knockout (-/-) and RLX wild-type (+/+) mice at various ages were analyzed for changes in collagen expression and content. We demonstrate an age-related progression of lung fibrosis in RLX -/- mice with significantly increased tissue wet weight, collagen content and concentration, alveolar congestion, and bronchiole epithelium thickening. The increased fibrosis was associated with significantly altered peak expiratory flow and lung recoil (lung function) in RLX -/- mice. Treatment of RLX -/- mice with relaxin in early and developed stages of fibrosis resulted in the reversal of collagen deposition. Organ bath studies showed that precontracted lung strips relaxed in the presence of relaxin. Together, these data indicate that relaxin may provide a means to regulate excessive collagen deposition in diseased states characterized by pulmonary fibrosis.