Comprehensive genomic analysis of sporadic multiple meningiomas reveals clonal origin and histotype-specific evolution: a case report

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Data on the genomic and molecular changes in sporadic multiple meningiomas are scarce, leading to ongoing debates regarding their evolutionary processes. A comprehensive genetic analysis of a large number of lesions, including minute occult meningiomas (MOMs), is necessary to explore these two possible origins: clonal and independent. In the present study, we performed whole-exome sequencing and analyzed somatic single-nucleotide variants (SNVs), insertions/deletions (INDELs), and copy number alterations (CNAs) in a patient with sporadic multiple meningiomas. These meningiomas included two mass-forming lesions of different histological subtypes (transitional and chordoid) and three MOMs. Genetic analysis revealed CNAs on chromosomes 22q and Y as common abnormalities in the two largest tumors. Furthermore, we identified SNV/INDELs unique to each focus, with NF2 mutation prevalent in the transitional meningioma and CREBBP mutation in the chordoid meningioma. Loss of chromosome 22 was detected in all three MOMs, whereas an NF2 somatic mutation was found only in the largest MOM. Overall, we elucidated the clonal origin and histotype-specific evolution of multiple meningiomas in this case. CNAs may serve as the initial driving event in meningioma development. multiple meningiomas occult meningioma copy number alteration whole-exome sequencing Figures Figure 1 Figure 2 Figure 3 Introduction Meningioma is the most common primary intracranial tumor in adults. Its incidence is higher in women than in men but the reason for this difference is unclear [ 1 ]. Most meningiomas are solitary; however, up to 10% of cases manifest as multiple tumors [ 2 ]. Recent evidence suggests that the incidence of multiple meningiomas may be even higher than 10% [ 3 ]. Multiple meningiomas are either sporadic or familial, with some sporadic cases being radiation-induced. The standard treatment for meningioma is surgical resection. In patients with multiple meningiomas, clinical decisions regarding the lesions that should be surgically resected and the order of their resection are often difficult. It is a chronic disease that requires repeated interventions and lifelong surveillance to achieve disease control [ 4 ]. In terms of prognosis, patients with multiple meningiomas exhibit shorter overall survival, progression-free survival, and time to second intervention than patients with a solitary meningioma [ 3 , 5 ]. Notably, a study that involved a large cohort of patients with multiple meningiomas revealed that a greater number of lesions, older age at diagnosis, and male sex were significantly negatively associated with overall survival [ 3 ]. Recent advances in the genomic analysis of solitary meningiomas have shed light on the relationships among the histological type, site of origin, malignancy, and prognosis [ 6 , 7 ]. Approximately 50% of meningiomas exhibit NF2 mutations and/or loss of chromosome 22, where NF2 is located. These genomic changes have been proven to be associated with atypical clinical and histological presentations due to genomic instability. Specifically, they show a predilection for meningiomas of the cerebrum, cerebellar hemispheres, posterior skull base, and spinal regions as well as transitional and fibrous subtypes [ 8 , 9 ]. Meningiomas without NF2 alterations are clinically benign and typically localized to the medial skull base. Their characteristic genomic changes include mutations of TRAF7 , KLF4 , AKT1 , and SMO . These mutations occur in a histotype-specific manner. Meningothelial and transitional meningiomas frequently harbor TRAF7 and either AKT1 or SMO mutations [ 9 ]. Mutations in SMARCE1 , BAP1 , or a combination of TRAF7 and KLF4 are associated with clear cell, rhabdoid, or secretory meningioma variants, respectively [ 9 , 10 ]. Multiple meningiomas are associated with familial tumor syndromes such as neurofibromatosis type 2 and schwannomatosis, which are genetically characterized by germline mutations of NF2 and SMARCB1 , respectively [ 11 ]. However, data on the genomic and molecular changes in patients with sporadic multiple meningiomas are scarce, leading to ongoing debates regarding their evolutionary processes [ 12 , 13 ]. Two hypotheses have been proposed to explain the pathogenesis of sporadic multiple meningiomas: a clonal origin and an independent origin. Studies supporting the clonality hypothesis suggest that multiple meningiomas arise from a specific neoplastic clone that proliferates along the meninges to form multifocal lesions [ 12 ]. This hypothesis is supported by observations that most sporadic multiple meningiomas exhibit identical histological features. By contrast, some researchers consider multiple meningiomas as independent lesions because some of these tumors exhibit various histological subtypes or grades [ 14 , 15 ]. A comprehensive genetic analysis of a large number of lesions, including minute occult meningiomas (MOMs), is necessary to resolve this issue. Although gaining a thorough understanding of meningiomas development requires the examination of MOMs, a putative precursor of mass-forming meningioma, no reports to date have discussed the genomic changes in MOMs. In the present study, we performed whole-exome sequencing (WES) and analyzed somatic single-nucleotide variants (SNVs), insertions/deletions (INDELs), and copy number alterations (CNAs) in a patient with sporadic multiple meningiomas. The meningiomas comprised two mass-forming lesions of different histological subtypes (transitional and chordoid) and three MOMs. The clonality and evolutional processes of these lesions were analyzed to elucidate the pathogenesis of sporadic multiple meningiomas. Clinical summary An 83-year-old man was incidentally discovered to have bilateral frontal convexity tumors 9 years prior to presentation. These asymptomatic tumors were monitored over time and identified as meningiomas through imaging. The patient had no familial history of the disease and no evidence of neurofibromatosis. He subsequently developed cognitive dysfunction coinciding with the identification of a new lesion on the left sphenoidal ridge. Preoperative magnetic resonance (MR) imaging revealed that this was the largest tumor, measuring 50 mm, and it exhibited strong homogenous enhancement. In addition, multiple smaller tumors up to 24 mm in size were observed in the bilateral convexity regions (Fig. 1 a). Pathological and genetic findings We simultaneously resected two largest mass-forming meningiomas, one in the left convexity (T1) and one on the left sphenoidal ridge (T2), along with the left convexity dura, which contained three MOMs (MOM1–MOM3). T1 was a transitional meningioma composed of proliferative meningothelial cells arranged in bundles or whorls (Fig. 1 b). T2 was a chordoid meningioma consisting of cord-like arrays of epithelioid cells within an abundant basophilic myxoid matrix (Fig. 1 c). T1 and T2 were grade 1 and 2 tumors, respectively, according to the 2016 World Health Organization (WHO) classification. In the dura surrounding the left convexity tumor, scattered MOMs composed of oval or spindle-shaped meningothelial cells with or without psammoma bodies were observed (Fig. 1 d–g). Immunohistochemistry showed that T1 and T2 were focally positive for epithelial membrane antigen (EMA) and progesterone receptor (PgR) and that T2 was negative for brachyury. The primary antibodies used in the immunohistochemical analysis are listed in Supplementary Table 1. WES was conducted to assess the clonality of the two resected meningiomas. Genomic DNA was extracted from frozen specimens of T1, T2 and background normal brain tissue. Libraries were prepared for each sample using a SureSelect Human All Exon V6 kit (Agilent Technologies) in accordance with the manufacturer’s recommendations. CNA analysis indicated loss of chromosomes 22q and Y in T1 and loss of chromosomes 1p, 10q, 22q, and Y in T2 (Fig. 2 a). Thus, loss of chromosomes 22q and Y were common events in both T1 and T2. Mutational analysis revealed 52 and 66 SNVs/INDELs in T1 and T2, respectively. After the application of strict filtering criteria, 14 mutations were retained in each tumor, with no overlapping mutations between T1 and T2 (Table 1 ). Among these genes, NF2 frameshift mutation (c.503delC:p.K170Rfs*43) in T1 and CREBBP frameshift mutation (c.3923delT:p.L1308Cfs*30) in T2 were highlighted upon comparison with previously published data on genes mutated in at least two cases of meningiomas (Fig. 2 b) [ 16 ]. Sanger sequencing was performed to validate the identified gene mutations, confirming the mutations of NF2 in T1 and CREBBP in T2 (Fig. 3 a). Homozygous deletions of CDKN2A and CDKN2B , as well as TERT promoter hotspot mutations that are indicative of WHO grade 3 tumors, were not detected in either T1 or T2 by WES and Sanger sequencing (data not shown). Table 1 Nonsynonymous gene mutations from the transitional meningioma (T1) and chordoid meningioma (T2) detected by whole-exome sequencing * Gene Mutation type Nucleotide change Amino acid change Allele frequency Transitional meningioma (T1) REPS2 Missense c.G650A p.S217N 74.3% NF2 Frameshift c.503delC p.K170Rfs*43 44.8% OXCT1 Missense c.G296A p.R99Q 43.8% ZNF469 Missense c.C11599G p.Q3867E 40.7% COL4A2 Missense c.C3076T p.P1026S 38.5% GLI4 Missense c.G283A p.G95R 38.4% TRPA1 Missense c.T1448C p.M483T 38.1% SOS1 Missense c.C3589G p.P1197A 34.7% PPP1R7 Missense c.G907C p.A303P 34.5% CLCN2 Missense c.C2096T p.S699F 34.5% DNAH5 Stop-gain c.C6763T p.R2255X 34.2% COL16A1 Missense c.C162G p.154M 33.1% CYB5R4 Missense c.G923T p.G308V 24.4% DNAH12 Missense c.A2274C p.K758N 20.8% Chordoid meningioma (T2) AHDC1 Missense c.G1760A p.R587Q 57.1% TYK2 Missense c.C2315T p.P1985fs 50.5% TNXB Frameshift c.5955delC p.P1985fs 48.0% SUCO Missense c.G530A p.S177N 44.5% DPH7 Missense c.C349T p.R117W 44.3% SUN1 Stop-gain c.G1638A p.W546X 44.2% NLRC3 Missense c.C1340T p.S447L 43.5% ZNF280D Missense c.C359T p.S120L 42.7% SLMAP Stop-gain c.C61T p.Q21X 41.9% NCOA6 Missense c.T3095C p.1032A 40.4% MYH7 Missense c.A3536G p.E1179G 39.8% CREBBP Frameshift c.3923delT p.L1308Cfc*30 37.3% TLR9 Missense c.A2432G p.D811G 30.6% RNF208 Stop-gain c.G198A p.V22M 29.5% * The filtering steps were performed according to the following four parameters: gnomad_AF_popmax < 0.01, normal VAF (NVAF) 0.2, and TVAF/NVAF > 5.0. Finally, to elucidate the clonal origin and histotype-specific evolution of multiple meningiomas, Sanger sequencing and Fluorescence in situ hybridization (FISH) of the three MOMs were performed, and the gene mutations and CNAs were compared to those of T1 and T2. The three MOMs of the dura mater were microdissected from sections stained with HE for DNA extraction. Sanger sequencing was performed to evaluate the gene mutations. Notably, sequencing of the three MOMs showed the same NF2 mutation in the largest MOM3 (Fig. 3 a), whereas the CREBBP mutation was not found in any of the specimens. The primers used in the Sanger sequencing are listed in Supplementary Table 2. Furthermore, FISH was performed to evaluate the copy number of chromosome 22 in the three MOMs using an NF2 (22q12) deletion probe (Guang Zhou LBP Medicine Science & Technology, Guangzhou, China). Each lesion was compared to a section stained with hematoxylin and eosin (HE) to aid the identification of proliferative meningeal cells. After identifying an internal control, such as endothelial cells or lymphocytes, the number of nuclei was counted in at least 30 cells. The criteria for determining the presence of loss of chromosome 22 was defined as the presence of ≥ 4.2% deleted cells based on a previous study using age- and sex-matched normal control bone marrow samples [ 17 ]. FISH of chromosome 22 in the three MOMs revealed scattered cells exhibiting monosomy of chromosome 22q with one red and one green signal (Fig. 3 b). The percentages of the meningeal cells displaying deletions in each lesion were as follows: 67.1% in MOM1, 40.9% in MOM2, and 50.0% in MOM3. All of these lesions were determined to have a loss of chromosome 22q. Collectively, they were considered to represent the clonal origin with loss of chromosome 22q, which then underwent histotype-specific evolution and acquired NF2 and CREBBP mutations (Fig. 3 c, d). Discussion We performed WES of solitary multiple meningiomas of different histological subtypes and identified CNAs on chromosomes 22q and Y as common genetic abnormalities. SNVs/INDELs unique to each focus were also detected. All of the lesions, including MOMs, were considered to be associated with NF2 based on the loss of chromosome 22q. Interestingly, the transitional meningioma (T1) had a second hit of NF2 and the chordoid meningioma (T2) had a CREBBP mutation. CREBBP is a chromatin-remodeling gene that is more enriched in chordoid than non-chordoid meningiomas [ 18 ]. This difference indicates that an epigenetic abnormality caused by CREBBP mutation may induce chordoid change against a background of chromosome 22 loss. The multiple meningiomas in this study were thought to be of clonal origin with histotype-specific evolution. A WES analysis of multiple meningiomas demonstrated clonal origin in five of six cases and subsequent branched evolution that resulted in inter-tumoral heterogeneity represented by different histologic types and grades [ 13 ]. Other studies of multiple meningiomas of different histological types have suggested that each lesion develops independently, but these studies did not evaluate CNAs and may not have accurately determined clonality [ 15 , 19 ]. Accurate assessment of the clonality of multiple meningiomas requires not only mutational analysis but also CNA analysis. Because multiple meningiomas may be both clonal and independent in origin, a large study is necessary. Our study is the first to analyze genetic changes in MOMs. Loss of chromosome 22 was detected in all three MOMs, whereas NF2 somatic mutation was found only in the largest MOM. It is often difficult to determine whether microscopic proliferative lesions consisting of meningothelial cells are reactive or neoplastic simply by observing their morphology. The presence of the same CNA in our patient indicated that the MOMs were neoplastic. The presence of CNAs has been reported in precancerous lesions such as intestinal metaplasia (which is a risk factor for gastric cancer) and clonal hematopoiesis (which is implicated in the development of hematological malignancies) [ 20 , 21 ]. Our findings indicate that CNAs are linked to the development of meningioma and that they precede SNVs/INDELs as genetic abnormalities, in line with a previous study of multiple meningiomas [ 13 ]. Chromosome 22q deletion is identified in approximately 50% of meningiomas, marking this as a critical early event in the onset of NF2 -related meningiomas [ 22 , 23 ]. Our study suggests that loss of chromosome Y (LOY) is also an initial event in the development of meningioma, concurrent with the loss of chromosome 22. However, the clinical and biological significance of the Y chromosome in meningioma remains largely unexplored. To date, only a few studies that have used FISH analysis of meningiomas have shown that LOY in men represents the second most frequent aberration, accounting for 28–46% of cases, following loss of chromosome 22 [ 17 , 24 ]. Qi et al . compiled an extensive catalog of LOY across more than 5,000 primary tumors from men in The Cancer Genome Atlas, demonstrating that LOY is exceedingly prevalent in numerous tumor types and suggesting its potential driving role in uveal melanoma [ 25 ]. Furthermore, LOY is associated with adverse outcomes in patients with bladder cancer, and cancer cells exhibiting LOY have been shown to modify T-cell functionality, leading to exhaustion of CD8 + T cells in the tumor microenvironment and increasing their susceptibility to PD-1-targeted immunotherapy [ 26 ]. In the context of meningioma, LOY may play a role in the tumorigenesis of meningiomas in men, potentially contributing to the poorer prognosis in men than in women. Conclusion Genomic analysis of solitary multiple meningiomas of different histological subtypes revealed the clonal origin and histotype-specific evolution. CNAs may serve as the initial driving event in meningioma development. Further investigation involving a larger cohort is warranted. Declarations Acknowledgement We thank Ms. Yuki Mitani for her excellent technical assistance. This work was supported by JSPS KAKENHI (Grant Number JP23H05340, MS). Conflict of interest The authors declare no conflict of interest. Ethical approval This study was approved by the Medical Ethics Committee of Kanazawa University (No.12644). Author Contributions Sakaguchi, Horie, Maeda contributed to the study conception and design. Material preparation was performed by Tanaka, Nakada. Data collection and analysis were performed by Sakaguchi, Horie, Ito, Mizuguchi, Ikeda, Kiyokawa, Maeda. The first draft of the manuscript was written by Sakaguchi, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Consent to participate Written informed consent was obtained from the patient. References Korhonen K, Salminen T, Raitanen J et al (2006) Female predominance in meningiomas can not be explained by differences in progesterone, estrogen, or androgen receptor expression. J Neurooncol 80(1):1–7 Tsermoulas G, Turel MK, Wilcox JT et al (2018) Management of multiple meningiomas. 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Cell Abdel-Hafiz HA, Schafer JM, Chen X et al (2023) Y chromosome loss in cancer drives growth by evasion of adaptive immunity. Nature 619(7970):624–631 Supplementary Files supplementary.docx Cite Share Download PDF Status: Published Journal Publication published 27 Jul, 2024 Read the published version in Brain Tumor Pathology → Version 1 posted Editorial decision: Major Revision 04 Jun, 2024 Reviewers agreed at journal 12 Apr, 2024 Reviewers invited by journal 12 Apr, 2024 Editor assigned by journal 29 Mar, 2024 First submitted to journal 28 Mar, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4183469","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":290378027,"identity":"d303683f-1026-4d89-9def-f849dd305106","order_by":0,"name":"Maki 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16:04:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4183469/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4183469/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10014-024-00486-9","type":"published","date":"2024-07-27T16:16:09+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":55002435,"identity":"b27c9b35-6ca5-4fe2-a25d-77eb46bb08e5","added_by":"auto","created_at":"2024-04-19 18:39:33","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1876143,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a) Gadolinium enhanced T1-weighted magnetic resonance image and (b–g) histological images of the patient in the present case\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn addition to the multiple lesions in the bilateral convexities, \u003cstrong\u003e(a)\u003c/strong\u003e a large tumor with strong homogenous gadolinium enhancement was present in the left sphenoidal ridge. The two large tumors located in the left convexity (T1) and on the left sphenoidal ridge (T2) were resected.\u003cstrong\u003e \u003c/strong\u003eArrowhead: non-resected multiple lesions in the convexity; arrow: resected tumors. Histologically, \u003cstrong\u003e(b)\u003c/strong\u003e T1 was composed of proliferative meningothelial cells arranged in bundles or whorls, whereas \u003cstrong\u003e(c)\u003c/strong\u003e T2 exhibited cord-like arrays of epithelioid tumor cells within an abundant basophilic myxoid matrix. \u003cstrong\u003e(d)\u003c/strong\u003e Macro-image of the three microdissected areas of the dura surrounding the left convexity tumor.\u003cstrong\u003e (e–g) \u003c/strong\u003eMicroscopically, three scattered minute occult meningiomas (MOMs) composed of oval or spindle-shaped meningothelial cells with or without psammoma bodies were identified.\u003c/p\u003e","description":"","filename":"Fig.1.png","url":"https://assets-eu.researchsquare.com/files/rs-4183469/v1/4ac1d7ef68904f24d9536acb.png"},{"id":55002438,"identity":"e25e1969-00a5-43f5-8eba-bc7f5a6ca4f6","added_by":"auto","created_at":"2024-04-19 18:39:34","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":461882,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a) Copy number alterations and (b) deletion mutations of T1 and T2 detected by whole-exome sequencing\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(a)\u003c/strong\u003e T1 showed loss of chromosomes 22q and Y, whereas T2 exhibited loss of chromosomes 1p, 10q, 22q, and Y. Homozygous deletions of \u003cem\u003eCDKN2A\u003c/em\u003e and \u003cem\u003eCDKN2B\u003c/em\u003e were not detected in either T1 or T2. \u003cstrong\u003e(b)\u003c/strong\u003e Information and coordinates of genetic mutations observed in T1 and T2. The \u003cem\u003eNF2\u003c/em\u003e frameshift mutation (c.503delC:p.K170Rfs*43) in T1 and\u003cem\u003e CREBBP\u003c/em\u003e frameshift mutation (c.3923delT:p.L1308Cfs*30) in T2 were considered significant.\u003c/p\u003e","description":"","filename":"Fig.2.png","url":"https://assets-eu.researchsquare.com/files/rs-4183469/v1/a7b35cbc5e95f8217f72186a.png"},{"id":55002436,"identity":"669d5175-3e82-4121-9aed-f49931f46d5f","added_by":"auto","created_at":"2024-04-19 18:39:33","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1817729,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMutation and copy number analyses of minute occults meningiomas (MOMs) by Sanger sequencing and fluorescence \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003ein situ\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e hybridization of chromosome 22\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(a)\u003c/strong\u003e The \u003cem\u003eNF2\u003c/em\u003e frameshift mutation in T1 and \u003cem\u003eCREBBP\u003c/em\u003e frameshift mutation in T2 were confirmed by Sanger sequencing. In addition, the same \u003cem\u003eNF2\u003c/em\u003emutation was detected in the largest MOM (MOM3). \u003cstrong\u003e(b)\u003c/strong\u003e All three MOMs (MOM1–MOM3) contained scattered cells exhibiting monosomy of chromosome 22q with one red and one green signal (arrowhead: heterozygous deletion; arrow: non-deleted). \u003cstrong\u003e(c)\u003c/strong\u003eSummary of CNA of chromosome 22 and mutations of \u003cem\u003eNF2\u003c/em\u003e and \u003cem\u003eCREBBP\u003c/em\u003e in three MOMs and two tumors. \u003cstrong\u003e(d)\u003c/strong\u003e Phylogeny inferred from the somatic CNA and SNV/INDEL.\u003c/p\u003e","description":"","filename":"Fig.3.png","url":"https://assets-eu.researchsquare.com/files/rs-4183469/v1/e2110548ad27a637d2cd1334.png"},{"id":61596197,"identity":"1d1cb003-8464-4786-9fbe-9c5a201d37d6","added_by":"auto","created_at":"2024-08-01 17:25:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":5652831,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4183469/v1/9bfd4f8d-f1d1-4184-9b3d-446c2d479aef.pdf"},{"id":55004784,"identity":"64eef0ec-0742-4b36-881a-c3d89fdfcdcd","added_by":"auto","created_at":"2024-04-19 18:47:33","extension":"docx","order_by":8,"title":"","display":"","copyAsset":false,"role":"supplement","size":22739,"visible":true,"origin":"","legend":"","description":"","filename":"supplementary.docx","url":"https://assets-eu.researchsquare.com/files/rs-4183469/v1/364518a2e5bdba8b6ba2703e.docx"}],"financialInterests":"","formattedTitle":"Comprehensive genomic analysis of sporadic multiple meningiomas reveals clonal origin and histotype-specific evolution: a case report","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMeningioma is the most common primary intracranial tumor in adults. Its incidence is higher in women than in men but the reason for this difference is unclear [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Most meningiomas are solitary; however, up to 10% of cases manifest as multiple tumors [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Recent evidence suggests that the incidence of multiple meningiomas may be even higher than 10% [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Multiple meningiomas are either sporadic or familial, with some sporadic cases being radiation-induced. The standard treatment for meningioma is surgical resection. In patients with multiple meningiomas, clinical decisions regarding the lesions that should be surgically resected and the order of their resection are often difficult. It is a chronic disease that requires repeated interventions and lifelong surveillance to achieve disease control [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. In terms of prognosis, patients with multiple meningiomas exhibit shorter overall survival, progression-free survival, and time to second intervention than patients with a solitary meningioma [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Notably, a study that involved a large cohort of patients with multiple meningiomas revealed that a greater number of lesions, older age at diagnosis, and male sex were significantly negatively associated with overall survival [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eRecent advances in the genomic analysis of solitary meningiomas have shed light on the relationships among the histological type, site of origin, malignancy, and prognosis [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Approximately 50% of meningiomas exhibit \u003cem\u003eNF2\u003c/em\u003e mutations and/or loss of chromosome 22, where \u003cem\u003eNF2\u003c/em\u003e is located. These genomic changes have been proven to be associated with atypical clinical and histological presentations due to genomic instability. Specifically, they show a predilection for meningiomas of the cerebrum, cerebellar hemispheres, posterior skull base, and spinal regions as well as transitional and fibrous subtypes [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Meningiomas without \u003cem\u003eNF2\u003c/em\u003e alterations are clinically benign and typically localized to the medial skull base. Their characteristic genomic changes include mutations of \u003cem\u003eTRAF7\u003c/em\u003e, \u003cem\u003eKLF4\u003c/em\u003e, \u003cem\u003eAKT1\u003c/em\u003e, and \u003cem\u003eSMO\u003c/em\u003e. These mutations occur in a histotype-specific manner. Meningothelial and transitional meningiomas frequently harbor \u003cem\u003eTRAF7\u003c/em\u003e and either \u003cem\u003eAKT1\u003c/em\u003e or \u003cem\u003eSMO\u003c/em\u003e mutations [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Mutations in \u003cem\u003eSMARCE1\u003c/em\u003e, \u003cem\u003eBAP1\u003c/em\u003e, or a combination of \u003cem\u003eTRAF7\u003c/em\u003e and \u003cem\u003eKLF4\u003c/em\u003e are associated with clear cell, rhabdoid, or secretory meningioma variants, respectively [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMultiple meningiomas are associated with familial tumor syndromes such as neurofibromatosis type 2 and schwannomatosis, which are genetically characterized by germline mutations of \u003cem\u003eNF2\u003c/em\u003e and \u003cem\u003eSMARCB1\u003c/em\u003e, respectively [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. However, data on the genomic and molecular changes in patients with sporadic multiple meningiomas are scarce, leading to ongoing debates regarding their evolutionary processes [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Two hypotheses have been proposed to explain the pathogenesis of sporadic multiple meningiomas: a clonal origin and an independent origin. Studies supporting the clonality hypothesis suggest that multiple meningiomas arise from a specific neoplastic clone that proliferates along the meninges to form multifocal lesions [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. This hypothesis is supported by observations that most sporadic multiple meningiomas exhibit identical histological features. By contrast, some researchers consider multiple meningiomas as independent lesions because some of these tumors exhibit various histological subtypes or grades [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. A comprehensive genetic analysis of a large number of lesions, including minute occult meningiomas (MOMs), is necessary to resolve this issue. Although gaining a thorough understanding of meningiomas development requires the examination of MOMs, a putative precursor of mass-forming meningioma, no reports to date have discussed the genomic changes in MOMs.\u003c/p\u003e \u003cp\u003eIn the present study, we performed whole-exome sequencing (WES) and analyzed somatic single-nucleotide variants (SNVs), insertions/deletions (INDELs), and copy number alterations (CNAs) in a patient with sporadic multiple meningiomas. The meningiomas comprised two mass-forming lesions of different histological subtypes (transitional and chordoid) and three MOMs. The clonality and evolutional processes of these lesions were analyzed to elucidate the pathogenesis of sporadic multiple meningiomas.\u003c/p\u003e"},{"header":"Clinical summary","content":"\u003cp\u003eAn 83-year-old man was incidentally discovered to have bilateral frontal convexity tumors 9 years prior to presentation. These asymptomatic tumors were monitored over time and identified as meningiomas through imaging. The patient had no familial history of the disease and no evidence of neurofibromatosis. He subsequently developed cognitive dysfunction coinciding with the identification of a new lesion on the left sphenoidal ridge. Preoperative magnetic resonance (MR) imaging revealed that this was the largest tumor, measuring 50 mm, and it exhibited strong homogenous enhancement. In addition, multiple smaller tumors up to 24 mm in size were observed in the bilateral convexity regions (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003ePathological and genetic findings\u003c/h2\u003e \u003cp\u003eWe simultaneously resected two largest mass-forming meningiomas, one in the left convexity (T1) and one on the left sphenoidal ridge (T2), along with the left convexity dura, which contained three MOMs (MOM1\u0026ndash;MOM3). T1 was a transitional meningioma composed of proliferative meningothelial cells arranged in bundles or whorls (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb). T2 was a chordoid meningioma consisting of cord-like arrays of epithelioid cells within an abundant basophilic myxoid matrix (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec). T1 and T2 were grade 1 and 2 tumors, respectively, according to the 2016 World Health Organization (WHO) classification. In the dura surrounding the left convexity tumor, scattered MOMs composed of oval or spindle-shaped meningothelial cells with or without psammoma bodies were observed (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ed\u0026ndash;g). Immunohistochemistry showed that T1 and T2 were focally positive for epithelial membrane antigen (EMA) and progesterone receptor (PgR) and that T2 was negative for brachyury. The primary antibodies used in the immunohistochemical analysis are listed in Supplementary Table\u0026nbsp;1.\u003c/p\u003e \u003cp\u003eWES was conducted to assess the clonality of the two resected meningiomas. Genomic DNA was extracted from frozen specimens of T1, T2 and background normal brain tissue. Libraries were prepared for each sample using a SureSelect Human All Exon V6 kit (Agilent Technologies) in accordance with the manufacturer\u0026rsquo;s recommendations. CNA analysis indicated loss of chromosomes 22q and Y in T1 and loss of chromosomes 1p, 10q, 22q, and Y in T2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea). Thus, loss of chromosomes 22q and Y were common events in both T1 and T2. Mutational analysis revealed 52 and 66 SNVs/INDELs in T1 and T2, respectively. After the application of strict filtering criteria, 14 mutations were retained in each tumor, with no overlapping mutations between T1 and T2 (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Among these genes, \u003cem\u003eNF2\u003c/em\u003e frameshift mutation (c.503delC:p.K170Rfs*43) in T1 and \u003cem\u003eCREBBP\u003c/em\u003e frameshift mutation (c.3923delT:p.L1308Cfs*30) in T2 were highlighted upon comparison with previously published data on genes mutated in at least two cases of meningiomas (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb) [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Sanger sequencing was performed to validate the identified gene mutations, confirming the mutations of \u003cem\u003eNF2\u003c/em\u003e in T1 and \u003cem\u003eCREBBP\u003c/em\u003e in T2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). Homozygous deletions of \u003cem\u003eCDKN2A\u003c/em\u003e and \u003cem\u003eCDKN2B\u003c/em\u003e, as well as \u003cem\u003eTERT\u003c/em\u003e promoter hotspot mutations that are indicative of WHO grade 3 tumors, were not detected in either T1 or T2 by WES and Sanger sequencing (data not shown).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eNonsynonymous gene mutations from the transitional meningioma (T1) and chordoid meningioma (T2) detected by whole-exome sequencing\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGene\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMutation type\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNucleotide change\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAmino acid change\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAllele frequency\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003eTransitional meningioma (T1)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eREPS2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G650A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.S217N\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.3%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eNF2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eFrameshift\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003ec.503delC\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003ep.K170Rfs*43\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e44.8%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eOXCT1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G296A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.R99Q\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e43.8%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eZNF469\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C11599G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.Q3867E\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.7%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCOL4A2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C3076T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.P1026S\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eGLI4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G283A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.G95R\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.4%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eTRPA1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.T1448C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.M483T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.1%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSOS1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C3589G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.P1197A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34.7%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePPP1R7\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G907C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.A303P\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCLCN2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C2096T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.S699F\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eDNAH5\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStop-gain\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C6763T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.R2255X\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e34.2%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCOL16A1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C162G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.154M\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e33.1%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCYB5R4\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G923T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.G308V\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e24.4%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eDNAH12\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.A2274C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.K758N\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20.8%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"5\" nameend=\"c5\" namest=\"c1\"\u003e \u003cp\u003eChordoid meningioma (T2)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAHDC1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G1760A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.R587Q\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e57.1%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eTYK2\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C2315T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.P1985fs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eTNXB\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFrameshift\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.5955delC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.P1985fs\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e48.0%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSUCO\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G530A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.S177N\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eDPH7\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C349T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.R117W\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.3%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSUN1\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStop-gain\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G1638A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.W546X\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.2%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eNLRC3\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C1340T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.S447L\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e43.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eZNF280D\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C359T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.S120L\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e42.7%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSLMAP\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStop-gain\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.C61T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.Q21X\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.9%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eNCOA6\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.T3095C\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.1032A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.4%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eMYH7\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.A3536G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.E1179G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e39.8%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCREBBP\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eFrameshift\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003ec.3923delT\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003ep.L1308Cfc*30\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e37.3%\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eTLR9\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMissense\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.A2432G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.D811G\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e30.6%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eRNF208\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eStop-gain\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ec.G198A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep.V22M\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e29.5%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003csup\u003e*\u003c/sup\u003e The filtering steps were performed according to the following four parameters: gnomad_AF_popmax\u0026thinsp;\u0026lt;\u0026thinsp;0.01, normal VAF (NVAF)\u0026thinsp;\u0026lt;\u0026thinsp;0.05, tumor VAF (TVAF)\u0026thinsp;\u0026gt;\u0026thinsp;0.2, and TVAF/NVAF\u0026thinsp;\u0026gt;\u0026thinsp;5.0.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eFinally, to elucidate the clonal origin and histotype-specific evolution of multiple meningiomas, Sanger sequencing and Fluorescence \u003cem\u003ein situ\u003c/em\u003e hybridization (FISH) of the three MOMs were performed, and the gene mutations and CNAs were compared to those of T1 and T2. The three MOMs of the dura mater were microdissected from sections stained with HE for DNA extraction. Sanger sequencing was performed to evaluate the gene mutations. Notably, sequencing of the three MOMs showed the same \u003cem\u003eNF2\u003c/em\u003e mutation in the largest MOM3 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea), whereas the \u003cem\u003eCREBBP\u003c/em\u003e mutation was not found in any of the specimens. The primers used in the Sanger sequencing are listed in Supplementary Table\u0026nbsp;2. Furthermore, FISH was performed to evaluate the copy number of chromosome 22 in the three MOMs using an \u003cem\u003eNF2\u003c/em\u003e (22q12) deletion probe (Guang Zhou LBP Medicine Science \u0026amp; Technology, Guangzhou, China). Each lesion was compared to a section stained with hematoxylin and eosin (HE) to aid the identification of proliferative meningeal cells. After identifying an internal control, such as endothelial cells or lymphocytes, the number of nuclei was counted in at least 30 cells. The criteria for determining the presence of loss of chromosome 22 was defined as the presence of \u0026ge;\u0026thinsp;4.2% deleted cells based on a previous study using age- and sex-matched normal control bone marrow samples [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. FISH of chromosome 22 in the three MOMs revealed scattered cells exhibiting monosomy of chromosome 22q with one red and one green signal (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). The percentages of the meningeal cells displaying deletions in each lesion were as follows: 67.1% in MOM1, 40.9% in MOM2, and 50.0% in MOM3. All of these lesions were determined to have a loss of chromosome 22q. Collectively, they were considered to represent the clonal origin with loss of chromosome 22q, which then underwent histotype-specific evolution and acquired \u003cem\u003eNF2\u003c/em\u003e and \u003cem\u003eCREBBP\u003c/em\u003e mutations (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec, d).\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eWe performed WES of solitary multiple meningiomas of different histological subtypes and identified CNAs on chromosomes 22q and Y as common genetic abnormalities. SNVs/INDELs unique to each focus were also detected. All of the lesions, including MOMs, were considered to be associated with \u003cem\u003eNF2\u003c/em\u003e based on the loss of chromosome 22q. Interestingly, the transitional meningioma (T1) had a second hit of \u003cem\u003eNF2\u003c/em\u003e and the chordoid meningioma (T2) had a \u003cem\u003eCREBBP\u003c/em\u003e mutation. \u003cem\u003eCREBBP\u003c/em\u003e is a chromatin-remodeling gene that is more enriched in chordoid than non-chordoid meningiomas [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. This difference indicates that an epigenetic abnormality caused by \u003cem\u003eCREBBP\u003c/em\u003e mutation may induce chordoid change against a background of chromosome 22 loss. The multiple meningiomas in this study were thought to be of clonal origin with histotype-specific evolution. A WES analysis of multiple meningiomas demonstrated clonal origin in five of six cases and subsequent branched evolution that resulted in inter-tumoral heterogeneity represented by different histologic types and grades [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Other studies of multiple meningiomas of different histological types have suggested that each lesion develops independently, but these studies did not evaluate CNAs and may not have accurately determined clonality [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Accurate assessment of the clonality of multiple meningiomas requires not only mutational analysis but also CNA analysis. Because multiple meningiomas may be both clonal and independent in origin, a large study is necessary.\u003c/p\u003e \u003cp\u003eOur study is the first to analyze genetic changes in MOMs. Loss of chromosome 22 was detected in all three MOMs, whereas \u003cem\u003eNF2\u003c/em\u003e somatic mutation was found only in the largest MOM. It is often difficult to determine whether microscopic proliferative lesions consisting of meningothelial cells are reactive or neoplastic simply by observing their morphology. The presence of the same CNA in our patient indicated that the MOMs were neoplastic. The presence of CNAs has been reported in precancerous lesions such as intestinal metaplasia (which is a risk factor for gastric cancer) and clonal hematopoiesis (which is implicated in the development of hematological malignancies) [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Our findings indicate that CNAs are linked to the development of meningioma and that they precede SNVs/INDELs as genetic abnormalities, in line with a previous study of multiple meningiomas [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChromosome 22q deletion is identified in approximately 50% of meningiomas, marking this as a critical early event in the onset of \u003cem\u003eNF2\u003c/em\u003e-related meningiomas [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. Our study suggests that loss of chromosome Y (LOY) is also an initial event in the development of meningioma, concurrent with the loss of chromosome 22. However, the clinical and biological significance of the Y chromosome in meningioma remains largely unexplored. To date, only a few studies that have used FISH analysis of meningiomas have shown that LOY in men represents the second most frequent aberration, accounting for 28\u0026ndash;46% of cases, following loss of chromosome 22 [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Qi \u003cem\u003eet al\u003c/em\u003e. compiled an extensive catalog of LOY across more than 5,000 primary tumors from men in The Cancer Genome Atlas, demonstrating that LOY is exceedingly prevalent in numerous tumor types and suggesting its potential driving role in uveal melanoma [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Furthermore, LOY is associated with adverse outcomes in patients with bladder cancer, and cancer cells exhibiting LOY have been shown to modify T-cell functionality, leading to exhaustion of CD8\u0026thinsp;+\u0026thinsp;T cells in the tumor microenvironment and increasing their susceptibility to PD-1-targeted immunotherapy [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In the context of meningioma, LOY may play a role in the tumorigenesis of meningiomas in men, potentially contributing to the poorer prognosis in men than in women.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eGenomic analysis of solitary multiple meningiomas of different histological subtypes revealed the clonal origin and histotype-specific evolution. CNAs may serve as the initial driving event in meningioma development. Further investigation involving a larger cohort is warranted.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Ms. Yuki Mitani for her excellent technical assistance.\u0026nbsp;This work was supported by JSPS KAKENHI (Grant Number JP23H05340, MS).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was approved by the Medical Ethics Committee of Kanazawa University (No.12644).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSakaguchi, Horie, Maeda contributed to the study conception and design. Material preparation was performed by Tanaka, Nakada. Data collection and analysis were performed by Sakaguchi, Horie, Ito, Mizuguchi, Ikeda, Kiyokawa, Maeda. The first draft of the manuscript was written by Sakaguchi, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWritten informed consent was obtained from the patient.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eKorhonen K, Salminen T, Raitanen J et al (2006) Female predominance in meningiomas can not be explained by differences in progesterone, estrogen, or androgen receptor expression. J Neurooncol 80(1):1\u0026ndash;7\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTsermoulas G, Turel MK, Wilcox JT et al (2018) Management of multiple meningiomas. J Neurosurg 128(5):1403\u0026ndash;1409\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamos-Fresnedo A, Domingo RA, Vivas-Buitrago T et al (2020) Multiple meningiomas: does quantity matter? a population-based survival analysis with underlined age and sex differences. J Neurooncol 149(3):413\u0026ndash;420\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFahlstrom A, Dwivedi S, Drummond K (2023) Multiple meningiomas: Epidemiology, management, and outcomes. Neurooncol Adv 5(Suppl 1):i35\u0026ndash;i48\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamos-Fresnedo A, Domingo RA, Sanchez-Garavito JE et al (2021) The impact of multiple lesions on progression-free survival of meningiomas: a 10-year multicenter experience. J Neurosurg. :1\u0026ndash;9\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYoungblood MW, Duran D, Montejo JD et al (2019) Correlations between genomic subgroup and clinical features in a cohort of more than 3000 meningiomas. J Neurosurg. :1\u0026ndash;10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoudhury A, Magill ST, Eaton CD et al (2022) Meningioma DNA methylation groups identify biological drivers and therapeutic vulnerabilities. Nat Genet 54(5):649\u0026ndash;659\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKros J, de Greve K, van Tilborg A et al (2001) NF2 status of meningiomas is associated with tumour localization and histology. J Pathol 194(3):367\u0026ndash;372\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eYuzawa S, Nishihara H, Tanaka S (2016) Genetic landscape of meningioma. 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BMC Med Genomics 15(1):112\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKoh YC, Yoo H, Whang GC et al (2001) Multiple meningiomas of different pathological features: case report. J Clin Neurosci 8(Suppl 1):40\u0026ndash;43\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJuratli TA, Prilop I, Saalfeld FC et al (2021) Sporadic multiple meningiomas harbor distinct driver mutations. Acta Neuropathol Commun 9(1):8\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNassiri F, Liu J, Patil V et al (2021) A clinically applicable integrative molecular classification of meningiomas. Nature 597(7874):119\u0026ndash;125\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSayagues JM, Tabernero MD, Maillo A et al (2002) Incidence of numerical chromosome aberrations in meningioma tumors as revealed by fluorescence in situ hybridization using 10 chromosome-specific probes. Cytometry 50(3):153\u0026ndash;159\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGeorgescu MM, Nanda A, Li Y et al (2020) Mutation Status and Epithelial Differentiation Stratify Recurrence Risk in Chordoid Meningioma-A Multicenter Study with High Prognostic Relevance. Cancers (Basel). ;12(1)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSheng HS, Shen F, Zhang N et al (2019) Whole exome sequencing of multiple meningiomas with varying histopathological presentation in one patient revealed distinctive somatic mutation burden and independent clonal origins. Cancer Manag Res 11:4085\u0026ndash;4095\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKumagai K, Shimizu T, Takai A et al (2022) Expansion of Gastric Intestinal Metaplasia with Copy Number Aberrations Contributes to Field Cancerization. Cancer Res 82(9):1712\u0026ndash;1723\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaiki R, Momozawa Y, Nannya Y et al (2021) Combined landscape of single-nucleotide variants and copy number alterations in clonal hematopoiesis. Nat Med 27(7):1239\u0026ndash;1249\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLomas J, Bello MJ, Arjona D et al (2005) Genetic and epigenetic alteration of the NF2 gene in sporadic meningiomas. Genes Chromosomes Cancer 42(3):314\u0026ndash;319\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee S, Karas PJ, Hadley CC et al (2019) The Role of Merlin/NF2 Loss in Meningioma Biology. Cancers (Basel). ;11(11)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDomingues PH, Sousa P, Otero A et al (2014) Proposal for a new risk stratification classification for meningioma based on patient age, WHO tumor grade, size, localization, and karyotype. Neuro Oncol 16(5):735\u0026ndash;747\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eQi M, Pang J, Mitsiades I et al (2023) Loss of chromosome Y in primary tumors. Cell\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAbdel-Hafiz HA, Schafer JM, Chen X et al (2023) Y chromosome loss in cancer drives growth by evasion of adaptive immunity. Nature 619(7970):624\u0026ndash;631\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"brain-tumor-pathology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"btpa","sideBox":"Learn more about [Brain Tumor Pathology](http://link.springer.com/journal/10014)","snPcode":"10014","submissionUrl":"https://www.editorialmanager.com/btpa/default2.aspx","title":"Brain Tumor Pathology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"multiple meningiomas, occult meningioma, copy number alteration, whole-exome sequencing","lastPublishedDoi":"10.21203/rs.3.rs-4183469/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4183469/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMeningioma is the most common primary intracranial tumor in adults, with up to 10% manifesting as multiple tumors. Data on the genomic and molecular changes in sporadic multiple meningiomas are scarce, leading to ongoing debates regarding their evolutionary processes. A comprehensive genetic analysis of a large number of lesions, including minute occult meningiomas (MOMs), is necessary to explore these two possible origins: clonal and independent. In the present study, we performed whole-exome sequencing and analyzed somatic single-nucleotide variants (SNVs), insertions/deletions (INDELs), and copy number alterations (CNAs) in a patient with sporadic multiple meningiomas. These meningiomas included two mass-forming lesions of different histological subtypes (transitional and chordoid) and three MOMs. Genetic analysis revealed CNAs on chromosomes 22q and Y as common abnormalities in the two largest tumors. Furthermore, we identified SNV/INDELs unique to each focus, with \u003cem\u003eNF2\u003c/em\u003e mutation prevalent in the transitional meningioma and \u003cem\u003eCREBBP\u003c/em\u003e mutation in the chordoid meningioma. Loss of chromosome 22 was detected in all three MOMs, whereas an \u003cem\u003eNF2\u003c/em\u003e somatic mutation was found only in the largest MOM. Overall, we elucidated the clonal origin and histotype-specific evolution of multiple meningiomas in this case. CNAs may serve as the initial driving event in meningioma development.\u003c/p\u003e","manuscriptTitle":"Comprehensive genomic analysis of sporadic multiple meningiomas reveals clonal origin and histotype-specific evolution: a case report","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-19 18:39:29","doi":"10.21203/rs.3.rs-4183469/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major Revision","date":"2024-06-04T23:43:43+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-04-12T07:09:44+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-04-12T06:28:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-03-29T05:24:31+00:00","index":"","fulltext":""},{"type":"submitted","content":"Brain Tumor Pathology","date":"2024-03-28T12:03:27+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"brain-tumor-pathology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"btpa","sideBox":"Learn more about [Brain Tumor Pathology](http://link.springer.com/journal/10014)","snPcode":"10014","submissionUrl":"https://www.editorialmanager.com/btpa/default2.aspx","title":"Brain Tumor Pathology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"fda97235-e370-41a6-a1f2-812766152a36","owner":[],"postedDate":"April 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-08-01T17:08:17+00:00","versionOfRecord":{"articleIdentity":"rs-4183469","link":"https://doi.org/10.1007/s10014-024-00486-9","journal":{"identity":"brain-tumor-pathology","isVorOnly":false,"title":"Brain Tumor Pathology"},"publishedOn":"2024-07-27 16:16:09","publishedOnDateReadable":"July 27th, 2024"},"versionCreatedAt":"2024-04-19 18:39:29","video":"","vorDoi":"10.1007/s10014-024-00486-9","vorDoiUrl":"https://doi.org/10.1007/s10014-024-00486-9","workflowStages":[]},"version":"v1","identity":"rs-4183469","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4183469","identity":"rs-4183469","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}