Robo signaling regulates the production of cranial neural crest cells

Purpose: The strong association between BAP1 mutations and metastasizing Class 2 uveal melanoma (UM) suggests that epigenetic alterations may play a significant role in tumor progression. Thus, we characterized the impact of BAP1 loss on the DNA methylome in UM.Experimental Design: Global DNA methylation was analyzed in 47 Class 1 and 45 Class 2 primary UMs and in UM cells engineered to inducibly deplete BAP1. RNA-Seq was analyzed in 80 UM samples and engineered UM cells.Results: Hypermethylation on chromosome 3 correlated with downregulated gene expression at several loci, including 3p21, where BAP1 is located. Gene set analysis of hypermethylated and downregulated genes identified axon guidance and melanogenesis as deregulated pathways, with several of these genes located on chromosome 3. A novel hypermethylated site within the BAP1 locus was found in all Class 2 tumors, suggesting that BAP1 itself is epigenetically regulated. Highly differentially methylated probes were orthogonally validated using bisulfite sequencing, and they successfully distinguished Class 1 and Class 2 tumors in 100% of cases. In functional validation experiments, BAP1 knockdown in UM cells induced methylomic repatterning similar to UM tumors, enriched for genes involved in axon guidance, melanogenesis, and development.Conclusions: This study, coupled with previous work, suggests that the initial event in the divergence of Class 2 UM from Class 1 UM is loss of one copy of chromosome 3, followed by mutation of BAP1 on the remaining copy of chromosome 3, leading to the methylomic repatterning profile characteristic of Class 2 UMs.

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“…3; refs. [43][44][45]. The proteins encoded by these axon guidance and melanogenesis genes demonstrated extensive interactome connectivity (Fig.…”

Section: Functional Pathway Analysismentioning

confidence: 99%

BAP1 Loss Is Associated with DNA Methylomic Repatterning in Highly Aggressive Class 2 Uveal Melanomas

Field

Kuznetsov

Bussies

et al. 2019

Clinical Cancer Research

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“…In contrast to the skeletal defects in the vertebral column, craniofacial defects of NELL1-deficient mice indicate a deficiency of neural crest cell derivatives (6). Recently, Li et al (26) demonstrated that Robo1 and Robo2 double knockout mouse embryos show the defect of cranial osteogenesis because production of cranial neural crest cells is reduced. The overlapping phenotypes of NELL1-deficient mice and Robo1/2 double knockout mice suggest that they function in a common pathway.…”

Section: Discussionmentioning

confidence: 99%

“…Robo receptors and their Slit ligands form one of the most crucial ligand-receptor pairings among axon guidance molecules. However, Slit/Robo signaling not only functions in axon guidance but also in a variety of developmental events outside the nervous system (26). For example, Slit/Robo signaling is involved in many aspects of heart development, such as cardiac cell migration and alignment (27).…”

Section: Discussionmentioning

confidence: 99%

Robo2 contains a cryptic binding site for neural EGFL-like (NELL) protein 1/2

Yamamoto

Kashiwagi

Ishihara

et al. 2019

Journal of Biological Chemistry

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The signaling pathways that are mediated by Slit ligands and their Roundabout (Robo) family of receptors play multifunctional roles in the development of the nervous system and other organs. A recent study identified neural epidermal growth factor-like (NEL)-like 2 (NELL2) as a novel ligand for Robo3. In this study, we carried out a comprehensive analysis of the interaction between NELL1 and the Robo family of receptors and demonstrated that Robo2 contains a cryptic binding site for both NELL1 and NELL2. NELL1/2 binds to the first fibronectin type III (FNIII) domain of Robo2 but not to intact Robo2. Mutation analysis revealed that several amino acids within the first FNIII domain are critical for NELL1 binding to Robo2 but not to Robo1. The Robo2 deletion mutants without the fourth immunoglobulin domain and single amino acid substitution mutants that can influence the architecture of the ectodomain facilitated binding to NELL1/2. Acidic conditions increased the binding affinity of Robo2 for NELL1. These results suggest that Robo2 functions as a receptor for NELL1/2, particularly under circumstances where Robo2 undergoes proteolytic digestion. If this is not the case, conformational changes of the ectodomain of Robo2 may unmask the binding site for NELL1/2.

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“…[ 109 ] Slit‐3 axon guidance cues emitting from the skeleton are sensed by nerves through the receptors, Robo1. [ 148–152 ] Ephrin B2, an Eph receptor‐interacting protein which navigates nerve growth, can be identified from osteoclasts and osteoblasts. [ 153 ] Osteoblasts produce Netrin‐1, the receptor of which is well expressed in the nervous system.…”

Section: Possible Signals From Bone To Nerves Within Bonementioning

confidence: 99%

Crosstalk between Bone and Nerves within Bone

et al. 2021

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For the past two decades, the function of intrabony nerves on bone has been a subject of intense research, while the function of bone on intrabony nerves is still hidden in the corner. In the present review, the possible crosstalk between bone and intrabony peripheral nerves will be comprehensively analyzed. Peripheral nerves participate in bone development and repair via a host of signals generated through the secretion of neurotransmitters, neuropeptides, axon guidance factors and neurotrophins, with additional contribution from nerve‐resident cells. In return, bone contributes to this microenvironmental rendezvous by housing the nerves within its internal milieu to provide mechanical support and a protective shelf. A large ensemble of chemical, mechanical, and electrical cues works in harmony with bone marrow stromal cells in the regulation of intrabony nerves. The crosstalk between bone and nerves is not limited to the physiological state, but also involved in various bone diseases including osteoporosis, osteoarthritis, heterotopic ossification, psychological stress‐related bone abnormalities, and bone related tumors. This crosstalk may be harnessed in the design of tissue engineering scaffolds for repair of bone defects or be targeted for treatment of diseases related to bone and peripheral nerves.

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Robo signaling regulates the production of cranial neural crest cells

Cited by 11 publications

References 68 publications

BAP1 Loss Is Associated with DNA Methylomic Repatterning in Highly Aggressive Class 2 Uveal Melanomas

BAP1 Loss Is Associated with DNA Methylomic Repatterning in Highly Aggressive Class 2 Uveal Melanomas

Robo2 contains a cryptic binding site for neural EGFL-like (NELL) protein 1/2

Crosstalk between Bone and Nerves within Bone

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