Clinicopathological features of NTRK3 gene rearrangement papillary thyroid carcinoma / 中华病理学杂志

Objective: To investigate the clinicopathological features and differential diagnosis of NTRK3 gene rearrangement thyroid papillary carcinoma (PTC). Methods: The PTC cases without BRAF V600E mutation were collected at Fujian Provincial Hospital South Branch from January 2015 to January 2020. The cases of NTRK3 gene rearrangement PTC were examined using immunohistochemistry and fluorescence in situ hybridization (FISH). The clinical data, histopathological characteristics, immunohistochemical features and molecular pathological changes were retrospectively analyzed. Data from the TCGA PTC dataset and the literature were also studied. Results: A total of 3 PTC cases harboring NTRK3 gene rearrangement were confirmed. All the patients were female, aged from 26,49,34 years. Histologically, two of them demonstrated a multinodular growth pattern. Only one case showed prominent follicular growth pattern; the other two tumors showed a mixture of follicular, papillary and solid growth patterns. All tumors showed a typical PTC nuclear manifestation, with some nuclear pleomorphism, vacuolated foci and oncocytic features. The characteristic formation of glomeruloid follicular foci was present in two cases which also showed psammoma bodies, and tumoral capsular or angiolymphatic invasion. The background thyroid parenchyma showed chronic lymphocytic thyroiditis. Mitotic rates were low, and no cases had any tumor necrosis. The pan-TRK and TTF1 testing was both positive in 3 cases, while S-100 and mammaglobin were both negative in them. FISH studies confirmed the NTRK3 gene rearrangement in all 3 cases. Studies on the TCGA datasets and literature revealed similar findings. Conclusions: NTRK3 gene rearrangement PTC is rare. It may be easily misdiagnosed due to the lack of histological and clinicopathological characteristics. Molecular studies such as pan-TRK immunostaining, FISH and even next-generation sequencing are needed to confirm the diagnosis. Immunohistochemistry of pan-TRK performed in the PTC cases without BRAF V600E mutation can be used as a good rapid-screening tool. With the emergence of pan-cancer tyrosine receptor kinase inhibitors, proper diagnosis of these tumors can help determine appropriate treatments and improve their outcomes.

[Effect of a session resistance exercise on mRNA expression of NT-3 and TrkC proteins in soleus muscle of Wistar rats]

Background and Objective: Skeletal muscle expresses several neurotrophin and their receptors which providing the basis for neurotrophin signaling within the muscle compartments. This study was done to evaluate the effect of a session of resistance exercise on mRNA expression of NT-3 and TrkC proteins in soleus muscle of Wistar Rats

Methods: In this experimental study, 16 male Wistar rats were randomly allocated into exercise and control groups. The resistance training protocol consisted of climbing a 1-meter-long ladder, with a weight attached to a tail sleeve. Expressions of NT-4/5 and P75, quantitatively were measured using RT-PCR

Results: There was not any significant alteration in NT-3 mRNA in soleus muscle after resistance exercise. However, one session of resistance exercise significantly increased mRNA expression of TrkC [1.7 Folds] in soleus muscle [P<0.05]

Conclusion: Resistance exercise increases TrkC expression in soleuse muscle of wistar rats

ShcD interacts with TrkC through its PTB and SH2 domains / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the interaction between ShcD and TrkC and to reveal the molecular mechanism of the downstream signal transduction of TrkC.</p><p><b>METHODS</b>Yeast two-hybrid assay was used. The intracellular domains of TrkC and TrkC mutants were cloned into pAS2-1, and ShcD and its four domains (CH2, PTB, CH1, and SH2 domains) were cloned into pACT2 vector respectively. The constructs were separately cotransformed into yeast. beta-galactosidase activity was measured to detect their interactions. TrkC was cloned into pmRFP (carrying red fluorescent protein), and ShcD was cloned into pEGFP (carrying green fluorescent protein). pmRFP-TrkC and pEGFP-ShcD were co-transfected into 293T cells, and then the cells were fixed and subjected to confocal analysis to study their subcellular localization.</p><p><b>RESULTS</b>ShcD interacted with TrkC but not with kinase dead mutant TrkCM1(K572A). Both PTB and SH2 domains were capable of binding to TrkC, and PTB domain bound NPQY motif of TrkC. ShcD colocalized with TrkC throughout the cytoplasm and in the plasma membrane in 293T cells.</p><p><b>CONCLUSION</b>ShcD binds to TrkC in a kinase-activity-dependent manner through its PTB and SH2 domains.</p>

Dok6 promote neurite outgrowth of tropomyosin-related kinase C overexpressed PC12 cells in stimulation of neurotrophin-3 / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the role of adaptor protein Dok6 in neurite outgrowth in PC12 cells.</p><p><b>METHODS</b>Series of fusion clones were constructed by fusing different domains of Dok6 into mutant TrkC/Y516F. These constructs were transiently transfected into PC12 cells separately and the expression levels of fusion proteins were detected by Western blot. Neurite outgrowth in these PC12 cells was tested after stimulation of NT-3.</p><p><b>RESULTS</b>Each fusion clone was stably expressed in PC12 cells. The fusion clones that fused both TrkC/Y516F-Dok6 (PTB+C) and TrkC/Y516F-Dok6C rescued the loss of neurite outgrowth in PC12 cells resulting from the mutation in tyrosine 516, while fusion clones that fused with single TrkC/Y516F-Dok6PTB did not show such effect.</p><p><b>CONCLUSION</b>Dok6 can promote neurite outgrowth induced by NT-3 stimulation through its C-terminal in TrkC-positive PC12 cells.</p>

Decreased expression of neurotrophic tyrosine receptor kinase 3 is associated with the outflow tract defect of human tetralogy of Fallot / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The molecular mechanism of human tetralogy of Fallot (TOF) is incompletely defined. Animal models have suggested that neurotrophic tyrosine receptor kinase 3 (NTRK3) might be associated with the outflow tract defect, similar to that seen in human TOF, however, the expression pattern of NTRK3 in human TOF heart tissues has never been investigated.</p><p><b>METHODS</b>Quantitative real-time PCR (qRT-PCR) and immunohistochemistry were applied to detect NTRK3 mRNA and protein levels in right ventricular outflow tract tissue samples of TOF patients, ventricular septal defect (VSD) patients and normal control infants (n = 10 in each group).</p><p><b>RESULTS</b>qRT-PCR analysis indicated that NTRK3 mRNA levels were significantly decreased in the TOF group compared to the VSD group (0.024 +/- 0.003 vs 0.085 +/- 0.004, P = 0.022) and the normal control group (0.024 +/- 0.003 vs 0.091 +/- 0.002, P = 0.006). Quantitative immunohistochemical analysis showed that NTRK3 protein was mainly localized in the myocardium cytoplasm in all 3 groups. The immunoreactivity of NTRK3 protein was again significantly lower in the TOF group compared to the VSD group (1.42 +/- 0.62 vs 14.12 +/- 1.83, P = 0.023) and the control group (1.42 +/- 0.62 vs 16.25 +/- 2.31, P = 0.008). The expression of NTRK3 in the VSD group and in the control group showed no significant differences at both mRNA and protein levels.</p><p><b>CONCLUSIONS</b>Insufficient expression of NTRK3 is associated with the outflow tract defect of human tetralogy of Fallot and may contribute to the progression of this defect.</p>

Expressions of neurotrophin factor receptor in spiral ganglion cell of cisplatin-induced ototoxicity / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of high-affinity tyrosine kinase receptors TrkB, TrkC and the low-affinity neurotrophin receptor p75 in spiral ganglion cell (SGC) of cisplatin-induced ototoxicity.</p><p><b>METHODS</b>The 50 adult Wistar rats were divided randomly into 5 groups received intraperitoneal injection of cisplatin with vary dose. Control group was received equivalent volumes of saline. The group received 1 day intraperitoneal injection was cisplatin treated at a dose of 5 mg/kg and killed at next day. The group received 3 days was cisplatin treated for 3 days at same dose daily and then killed at next day. The group A received 5 days was cisplatin treated for 5 days and killed at next day. The group B received 5 days was cisplatin treated for 5 days and then were sacrificed after 7 days. The change of mRNA level of neurotrophin receptors in cochlear tissue were examined by RT-PCR. The expressing pattern of TrkB, TrkC, P75 in damaged cochlea were study by immunochemistry using antibodies against TrkB, TrkC, P75 protein.</p><p><b>RESULTS</b>The research data showed the expression of Trk B, Trk C, p75 exhibited in SGC was dynamic along with the administration lasting. The mRNA and protein level of Trk B (x(-) +/- s) at day 1 and 3 after cisplatin treatment were 0.76 +/- 0.06, 88.78 +/- 4.28, 0.82 +/- 0.09 and 91.64 +/- 4.06, with significant difference among those and other groups (P < 0.05). The mRNA and protein level of TrkC at day 1 after cisplatin treatment were 0.80 +/- 0.06 and 89.66 +/- 2.76, with significant difference among that and other groups (P < 0.05). The mRNA and protein level of p75 at the control group and cisplatin treated groups were 0.64 +/- 0.04, 55.16 +/- 3.10, 0.77 +/- 0.04, 78.46 +/- 3.86, 1.01 +/- 0.09, 105.02 +/- 6.61, 1.18 +/- 0.09, 111.10 +/- 6.08, 0.51 +/- 0.04 and 42.74 +/- 5.20, with significant difference among the control group and cisplatin treated groups (P < 0.05).</p><p><b>CONCLUSIONS</b>The expression of Trk B increased to peak at day 1 - 3 after cisplatin treatment and decreased at day 5 early and following weeks. The expression of Trk C went up to peak at day 1 after cisplatin treatment and went down during subsequently time. P75 kept a trend of continuance increased during the drug treatment and decrease at drug stopped. The expression of Trk B, Trk C and P75 may be involved in cochlear insult with cisplatin-induced. Trk B and Trk C may play an important role in the reparative process of cochlear, especially at early stage of the damage. P75 could promote SGC apoptosis in cisplatin-induced neurotoxicity.</p>

Immunohistochemical Expressions of TrkB and TrkC Receptors in Rat Cochleas with Amikacin-induced Ototoxicity / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Neurotrophins have been known to be responsible for the differentiation and survival of developing neurons as well as for aiding the recovery of adult neurons from injury. The neurotrophin family includes NGF, BDNF, NT-3, and NT-4/5, and they exert their biological functions through activation of the high-affinity binding receptors, that is trkA, trkB, and trkC, with high characteristic specificity. Previous studies indicate that spiral ganglion cells express trkB and trkC mRNAs, while auditory hair cells produce NT-3 mRNA that directly affect maturation and survival of auditory neurons. It has been reported that the loss of target innervation and the eventual degeneration of auditory neurons caused by aminoglycoside ototoxicity can be prevented by the infusion of neurotrophic factors. The purpose of this study is to provide the expression patterns of trkB and trkC in the normal chochleas and damaged cochleas with aminoglycoside ototoxicity. MATERIALS AND METHODS: Adult Sprague-Dawley rats were treated with amikacin 500 mg/kg for ten days, and sacrificed on the 7th, 14th, 21th, and 28th day following the last injection. Auditory brainstem response was measured in each animal. Immunohistochemical method was used to study the localization of trkB and trkC receptors in the cochleas of adult rats of either normal control group or ototoxicity group. RESULTS: Immunoreactivities to trkB and trkC receptors were strongly positive in the spiral ganglion cells of all cochleas, especially in the neuronal perikarya of the type I cells. No difference in staining pattern was seen among the cochleas with different hearing thresholds. CONCLUSION: The uniform expression pattern of trkB and trkC receptors in the spiral ganglion cells regardless of the degree of ototoxicity suggests that neurotrophic factors may bind to these receptors to initiate the cellular mechanisms for neuronal survival in the injured auditory system.