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Homeodomain Transcription Factors Nkx2.2 and Pax6 as Novel Biomarkers for Meningioma Tumor Treatment

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Abstract

Meningioma is a common brain tumour which has neither a specific detection nor treatment method. The Sonic hedgehog (Shh) cell signaling pathway is a crucial regulatory pathway of mammalian organogenesis and tumorigenesis including meningioma. Shh cell signalling pathway cascade function by main transcription factor Gli1 and which further regulates in its downstream to Pax6 and Nkx2.2. This current study is aimed to explore the regulation of the Sonic hedgehog-Gli1 cell signaling pathway and its potential downstream targets in meningioma samples. A total of 24 surgically resected meningioma samples were used in this current study.Cytological changes were assessed using electron microscopic techniques as well as hematoxylin & eosin and DAPI staining. The expression pattern of Gli1, Nkx2.2 and Pax6 transcription factors were determined by using immunohistochemistry. The mRNA expression was assessed using RT-qPCR assays. Later, the whole transcriptome analysis of samples was performed with the amploseq technique. Results were compared with those obtained in normal human brain tissue (or normal meninges). Compared to the normal human brain tissue, meningioma samples showed crowded nuclei with morphological changes. Transcription factor Nkx2.2 expressed highly in all samples (24/24, 100%). Twenty-one of the 24 meningiomas (88%) showed high Gli1 and Pax6 expression. Whole transcriptome analysis of two meningioma samples also exhibited a very high increase in Gli1 expression signal in meningioma samples as compare to normal control. Hence, we may conclude that the Shh–Gli1 pathway is aberrantly activated in meningioma cells and is canonically upregulating the expression of transcription factors Pax6 and Nkx2.2.

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Data Availability

The authors confirm the data available in the manuscript and supplementary material. Raw data could be available from the corresponding author on request.

Abbreviations

SHH:

Sonic Hedgehog

M:

Meningioma

Gli1:

Glioma associated oncogene

Pax6:

Paired Box 6

Nkx2.2:

NK2 Homeobox 2

Patch1:

Patched 1

SMO:

Smoothened

SuFu:

Suppressor of fused homolog

HGMs:

High-grade meningiomas

LOH:

Loss of heterozygosity

TMZ:

Temozolomide

WNT:

Wingless-related integration site

MAPK:

Mitogen-activated protein kinase

CNS:

Central nervous system

DPX:

Dibutylphthalate polystyrene xylene

DAPI:

4’,6-Diamidino-2-phenylindole

PFA:

Paraformaldehyde

PBSt:

Phosphate-buffered saline/tween

EtBr:

Ethidium bromide

RT-qPCR:

Reverse transcription quantitative real-time PCR

mRNA:

Messenger RNA

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Acknowledgements

We acknowledge all the Interdisciplinary Brain Research Centre (IBRC) members, the financial support received from the DBT(6242‐P 102/RGCB/PMD/DBT/MDHS/2015) Council for Scientific and industrial research (CSIR) and UGC, and the IBRC, Faculty of Medicine, J.N. Medical College, Aligarh Muslim University for providing a platform for performing research.

Funding

This study funded by D.B.T. (6242‐P 102/RGCB/PMD/DBT/MDHS/2015) Council for Scientific and Industrial Research (CSIR) 09/112 (0621) 2k19EMR-1.

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Authors and Affiliations

  1. Interdisciplinary Brain Research Centre, J. N. Medical College, Faculty of Medicine, Aligarh Muslim University (A.M.U), Aligarh, 202002, Uttra Pradesh, India

    Shirin Farheen, Mubeena Mariyath PM, Asif Ali & Mehdi H. Shahi

  2. Department of Pathology, J. N. Medical College, Faculty of Medicine, Aligarh Muslim University, Aligarh, India

    Suhailur Rehman

  3. Department of Neurosurgery, J. N. Medical College, Faculty of Medicine, Aligarh Muslim University, Aligarh, India

    Md. Fakhrul Hoda

  4. Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

    Yakhlesh Gupta & Kunzang Chosdol

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Contributions

The original manuscript was drafted by the first author SF. SF materials and method preparation, data collection and analysis of the manuscript. PMMM, the second author of this manuscript, has formal analysis and methodology. The third author of this manuscript, SR, provided the tumour samples' grades for the histopathology assessment. Fourth author, MFH contributed by offering resources; the meningioma samples were assessed for the study. YG helped in evaluating the real-time PCR technique. AA investigated and supervised throughout the study and contributed with a partial funding source for the experiments included in this article. KC has helped by contributing to project administration and data curation. Lastly, the manuscript's corresponding author, MHS, supervised the complete study and acquired funding from DBT and the ICMR Government of India.

Corresponding author

Correspondence to Mehdi H. Shahi.

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Authors declare no conflict of interest.

Ethical approval

The protocols performed in the study involving human samples aligned with the institutional ethical standards; or any practices conducted by the studies. The Institutional Ethics Committee meeting was held on 14–03-2018, with registration number D approving all the experimental protocols. No. 926/F.M.

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All the paper participants have received consent to publish.

Informed consent in the manuscript

This study received ethical approval from the Ethical Committee of Jawaharlal Nehru Medical College, Aligarh Muslim University and performed following the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards. All procedures performed in this study involving human clinical samples followed the ethical standards of the Institutional Ethics Committee meeting held on 14-03-2018 with registration number D. No. 926/F.M. and the 1964 Helsinki declaration and later amendments or comparable ethical standards. Patients have given written consent for this study with their thumb impressions and initials to use their excised tumor samples.

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Farheen, S., PM, M.M., Rehman, S. et al. Homeodomain Transcription Factors Nkx2.2 and Pax6 as Novel Biomarkers for Meningioma Tumor Treatment. Ind J Clin Biochem 39, 47–59 (2024). https://doi.org/10.1007/s12291-022-01085-1

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