Comparative Transcriptomic Analysis of Primary and Recurren…

Comparative Transcriptomic Analysis of Primary and Recurrent Ovarian Granulosa Cell Tumors Eleonora Khlebus a* , Veena K. Vuttaradhi a , Thomas Welte a , Namrata Khurana a , Joseph Celestino b , Hannah C. Beird a , Curtis Gumbs a , Latasha Little a , Tri Nguyen b , Barrett Lawson c , Russell R. Broaddus d , David M. Gershenson b , P. Andrew Futreal a , and R. Tyler Hillman a,b a Department of Genomic Medicine, b Department of Gynecologic Oncology and Reproductive Medicine, c Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA d Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, NC, 27599, USA *eykhlebus@mdanderson.org

Leading Edge of Cancer Research Symposium 2022

Cryopreserved tumor tissue samples (n=24)

Recurrent aGCTs (n=16)

Primary aGCTs (n=8)

vs

Granulosa cells

Oocyte

• Adult-type granulosa cell tumor (aGCT) is a rare ovarian sex-cord stromal tumor (~ 5% of all ovarian tumors) • Most primary aGCTs are diagnosed at an early stage and can be removed surgically with long-term survival rates exceeding 90% • aGCT recurrence is difficult to predict and is almost always incurable after relapse The objective: Identify the differences between Primary and Recurrent aGCTs which may correlate with recurrence

Bulk RNA-sequencing

Differential Gene Expression analysis

Tumor Microenvironment analysis

Ranked gene list

Differentially expressed genes

Gene Set Enrichment Analysis (GSEA)

Enriched gene sets

Eleonora Khlebus eykhlebus@mdanderson.org

Differential Gene Expression analysis: Recurrent vs Primary tumors

• Functional Enrichment analysis: Gene Set Enrichment

• 31 differentially expressed gene identified (P adj < 0.05, FoldChange > 2)

Higher Expression in Primary Tumors : • LHCGR and INSL3 - genes involved in normal ovarian sex cord hormonal signaling Higher Expression in Recurrent Tumors: • NELL2 , GDF6 , and IL1RL1 - in some cancer types increases expression of these genes were associated with increased metastasis and decreased survival rate • CYP19A1 (aromatase) expression may be closely linked to aGCT pathogenesis and progression

Primarily immune-related and hormone-regulated gene sets expression was increased in recurrent tumors

Eleonora Khlebus eykhlebus@mdanderson.org

Comparative tumor microenvironment analysis

Tumor microenvironment analysis

• Tumor microenvironment in silico cytometry

Higher fractions in Recurrent tumors Lower fractions in Recurrent tumors

CIBERSORTx

Modified from Newman et al. (2019) Nat Biotech.

Conclusions:

§ Tumor microenvironment composition is significantly different between primary and recurrent aGCTs • Recurrent tumors have increased myeloid fraction and reduced prevalence of stromal cells what can promote tumor progression § Expression of genes involved in ovarian hormone signaling was identified as significantly altered in recurrences • Changes in hormonal signaling pathways may underpin aGCT recurrence

Financial support: This work was supported by grants from the Cancer Prevention and Research Institute of Texas (CPRIT) to R.T.H. (RR200045) and P.A.F. (R1205). R.T.H. is supported by the Jennifer “Jenny” Song Fund for Granulosa Cell Tumor Research. The University of Texas MD Anderson Cancer Center Multidisciplinary Gynecologic Cancer Tumor Bank is supported in part by NIH P50CA83639 SPORE in Ovarian Cancer. The authors declare no potential conflicts of interest.

Eleonora Khlebus eykhlebus@mdanderson.org

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