Characteristic of Uterine Rhabdomyosarcoma by Algorithm of Potential Biomarkers for Uterine Mesenchymal Tumor
, Tomoyuki Ichimura 4, Nobuo Yaegashi 5, Kaoru Abiko 1 and Ikuo Konishi 1,6Round 1
Reviewer 1 Report
In general, the work is well explained and well written. The discovery of new biomarkers to distinguish different uterine tumours is very useful and could help in faster correct diagnosis and selection of the most appropriate treatment. Although it could be interesting if more case reports were included and compared. This would allow to determine whether the selected molecular biomarkers follow the same patterns in tumours derived from other patients.
On the other hand, an author self-cites at least 12 times, considering that the current paper has 28 citations, authors should avoid excessive self-citation.
Author Response
Manuscript ID: curroncol-1644828
Reply to revierwer's comments
Reviewer 1:
In general, the work is well explained and well written. The discovery of new biomarkers to distinguish different uterine tumours is very useful and could help in faster correct diagnosis and selection of the most appropriate treatment.
Comment 1. Although it could be interesting if more case reports were included and compared. This would allow to determine whether the selected molecular biomarkers follow the same patterns in tumours derived from other patients.
Answer 1. We appreciate the reviewer’s comment. We agree with Reviewer's comments. We considered the issues pointed out by the reviewer and added them in discussion section in the revised manuscript as shown below.
Sarcomas are rare and usually display no specific line of differentiation, although rhabdomyosarcoma and leiomyosarcoma have been described (24,25). Recent study shows that NT5DC2 is aberrantly upregulated in uterine leiomyosarcoma, the expression of cyclin B1, cyclin A2, cyclin E1 is depended on the expression of NT5DC2 (26). However, the expression status of NT5DC2 in various mesenchymal tumors including benign uterine leiomyoma has not been clarified. Therefore, it is not clear whether NT5DC2 is a candidate biomarker for differentiating uterine leiomyosarcoma or rhabdomyosarcoma from other mesenchymal tumors. As pathogenesis for uterine leiomyosarcoma, the most frequently mutated genes include TP53, ATRX, and MED12, however no specific pathogenic variant has been identified (27,28). The other hand, molecular studies of a single case of pleomorphic rhabdomyosarcoma revealed PIK3CA and TP53 mutations (29). Therefore, it is inappropriate to distinguish between uterine leiomyosarcoma and uterine rhabdomyosarcoma based on the analysis results of pathological variants.
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- Hu B, Zhou S, Hu X, Zhang H, Lan X, Li M, Wang Y, Hu Q. NT5DC2 promotes leiomyosarcoma tumour cell growth via stabilizing unpalmitoylated TEAD4 and generating a positive feedback loop. J Cell Mol Med. 2021 May 16;25(13):5976-87. doi: 10.1111/jcmm.16409.
- An Y, Wang S, Li S, Zhang L, Wang D, Wang H, Zhu S, Zhu W, Li Y, Chen W, Ji S, Guo X. Distinct molecular subtypes of uterine leiomyosarcoma respond differently to chemotherapy treatment. BMC Cancer. 2017 Sep 11;17(1):639. doi: 10.1186/s12885-017-3568-y.
- Ravegnini, G.; Mariño-Enriquez, A.; Slater, J.; Eilers, G.; Wang, Y.; Zhu, M.; Nucci, M.R.; George, S.; Angelini, S.; Raut, C.P.; Fletcher, J.A. MED12 mutations in leiomyosarcoma and extrauterine leiomyoma. Mod Pathol. 2013; 26(5): 743-749.
- Pinto A, Kahn RM, Rosenberg AE, Slomovitz B, Quick CM, Whisman MK, Huang M. Uterine rhabdomyosarcoma in adults. Hum Pathol. 2018 Apr;74:122-128. doi: 10.1016/j.humpath.2018.01.007.
Comment 2. On the other hand, an author self-cites at least 12 times, considering that the current paper has 28 citations, authors should avoid excessive self-citation.
Answer 2. We appreciate the reviewer’s comment. We agree with Reviewer's comments.
Following the reviewer's comments, we made corrections and additions throughout the manuscript. Finally, a total of 40 references are cited in the revised manuscript. Ten of our papers have been cited in the revised manuscript.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The manuscript entitled with "Characteristic of Uterine Rhabdomyosarcoma by algorithm of 2 potential biomarkers for uterine mesenchymal tumor" give a try to find a new molecular biomarkers for the diagnosis of gynecologic tumors.
I have following specific comments.
1) it will be easier for the readers if the basic sample information is listed in the method and material instead of in the supplementary.
2) more reference is needed for the introduction part, for example, from line 74 to 78, for those marker proteins, it will be better to put reference there.
3) line 173 "many tumor cells the uterine tumor were round" is confusing, not sure what the authors trying to tell.
Author Response
Manuscript ID: curroncol-1644828
Reply to revierwer's comments
Reviewer 2:
I have following specific comments.
Comment 1. it will be easier for the readers if the basic sample information is listed in the method and material instead of in the supplementary.
Answer 1. We appreciate the reviewer’s comment. We agree with Reviewer's comments. We considered the issues pointed out by the reviewer. We have added the content described in the supplement to the section of materials and materials in the manuscript.
Tissue Collection. A total of 101 patients between 32 and 83 years of age and diagnosed as having smooth muscle tumors of the uterus were selected from pathological files. Serial sections were cut from at least 2 tissue blocks from each patient for hematoxylin and eosin staining and immunostaining. All tissues were used with the approval of the Ethical Committee of Shinshu University after obtaining written consent from each patient. The pathological diagnosis of uterine smooth muscle tumors was performed using established criteria (Hendrickson and Kempson, 1995) with some modification. Briefly, usual leiomyoma (usual LMA) was defined as a tumor showing typical histological features with a mitotic index (MI) [obtained by counting the total number of mitotic figures (MFs) in 10 high-power fields (HPFs)] of <5 MFs per 10 HPFs. Cellular leiomyoma (cellular LMA) was defined as a tumor with significantly increased cellularity (>2000 myoma cells / HPF) and a MI<5, but without cytologic atypia. Bizarre leiomyoma (BL) was defined as a tumor either with diffuse nuclear atypia and a MI<2 or with focal nuclear atypia and a MI<5 without coagulative tumor cell necrosis. A tumor of uncertain malignant potential (UMP) was defined as tumor with no mild atypia and a MI<10 but with coagulative tumor cell necrosis. Leiomyosarcoma (LMS) was diagnosed in the presence of a MI>10 with either diffuse cytologic atypia, coagulative tumor cell necrosis, or both. Of the 105 smooth muscle tumors, 52 were diagnosed as LMA, 3 were BL, 2 were intravenous leiomyomatosis, 58 were uterine LMS, 1 was uterine LANT-like tumor, and 2 were uterine rhabdomyosarcoma. Of the 58 LMS, 48 were histologically of the spindle-cell type and 10 were of the epithelioid type. The clinical stage of the LMS patients was stage I in 11 cases, stage II or III in 31 cases, and stage IV in 16 cases. Protein expression studies with cervix epithelium and carcinoma tissues were performed using tissue array (Uterus cancer tissues, AccuMax Array, Seoul, Korea). Details about tissue sections are indicated in manufacture’s information (AccuMax Array).
Comment 2. more reference is needed for the introduction part, for example, from line 74 to 78, for those marker proteins, it will be better to put reference there.
Answer 2. We appreciate the reviewer’s comment. We agree with Reviewer's comments. Following the reviewer's instructions, we have added the appropriate references in the text as follows:
In many cases of uterine adenosarcoma, the expression of cluster of differentiation (CD) 10, an epithelial cell marker, is diffusely positive and the expression of alpha smooth muscle actin (aSMA) and desmin, a marker of smooth muscle cells, is negative (6,7). In addition, in cases of uterine adenosarcoma, the expression of the hormone receptors, estrogen receptor (ER) and progesterone receptor (PgR), may be positive or negative (8,9).
- 6. Nathenson MJ, Ravi V, Fleming N, Wang WL, Conley A. Uterine Adenosarcoma: a Review. Curr Oncol Rep. 2016 Nov;18(11):68. doi: 10.1007/s11912-016-0552-7.
- 7. Oliva E. Practical issues in uterine pathology from banal to bewildering: the remarkable spectrum of smooth muscle neoplasia. Mod Pathol. 2016 Jan;29 Suppl 1:S104-20. doi: 10.1038/modpathol.2015.139.
- 8. Soslow RA, Ali A, Oliva E. Mullerian adenosarcomas: an immunophenotypic analysis of 35 cases. Am J Surg Pathol. 2008 Jul;32(7):1013-21. doi: 10.1097/PAS.0b013e318161d1be.
- 9. Marcus JZ, Klobocista M, Karabakhtsian RG, Prossnitz E, Goldberg GL, Huang GS. Female Sex Hormone Receptor Profiling in Uterine Adenosarcomas. Int J Gynecol Cancer. 2018 Mar;28(3):500-504. doi: 10.1097/IGC.0000000000001183.
Comment 3. line 173 "many tumor cells the uterine tumor were round" is confusing, not sure what the authors trying to tell.
Answer 3. We appreciate the reviewer’s comment. We agree with Reviewer's comments. Following the reviewer's instructions, we have rewritten the text as follows.
Many tumor cells the uterine tumor were round, short-spindle-shaped cells with a high nuclear cytoplasmic ratio (N/C).
Many tumor cells found in a patient's uterine tumor are round-shaped cells or short-spindle-shaped cells with a high nuclear-cytoplasmic ratio (N/C).
Author Response File: Author Response.pdf
