A Patient-Derived Glioblastoma Organoid Model that Preserves Intra- and Intertumoral Heterogeneity

Abstract

Glioblastoma (GB) is an aggressive brain tumor characterized by extensive heterogeneity. We aimed to generate patient-derived GB organoids preserving histological features and to establish a biobank for future studies. Materials and Methods: Tumor tissues from GB patients were dissected into 0.5–1 mm fragments and cultured for 14 days in organoid medium under orbital shaking. Immunohistochemical analysis was performed to assess GFAP, OLIG2, IDH-1, p53, ATRX, and Ki67 expression before and after organoid formation. Results: All tumor samples formed spherical organoids within 14 days. Histological analysis showed preservation of GB-specific morphological features, including nuclear atypia and pleomorphism. GFAP, OLIG2, and ATRX expression patterns were concordant between parental tumors and organoids in cases. IDH-1 expression was consistent in three patients. Ki67 and p53 expression levels showed similar patterns between organoids and corresponding tumors. Conclusion: Patient-derived GB organoids retain key histopathological features and may serve as a platform for GB modeling.

Keywords

• Glioblastoma
• organoid culture
• tumor heterogeneityse

Supporting Institution

We greatly acknowledge to the Scientific and Technological Research Council of Turkey (TÜBİTAK) for supporting author Cagla Tekin within the scope of the 2211-A doctoral program scholarship.

Project Number

TAY-2022-584

Ethical Statement

Ethical approval for this study was granted by the Bursa Uludag University Ethics Committee (Dated 20.06.2021, decision no: 2021-9/14). All patients were fully informed about the study objectives, confidentiality measures, and their right to withdraw at any time, and written informed consent was obtained prior to participation. The study was conducted in full accordance with the Declaration of Helsinki.

Thanks

We thanks to Prof. Dr. Şükrü Yıldırım from Istanbul Maltepe University, Prof. Dr. Şahsine Tolunay and Assoc. Prof. Dr. Mine Ozsen from Bursa Uludag University for their support in IHC analysis. We would also like to thanks to Prof. Dr. Bahattin Hakyemez for special assistance with MRI imaging.

İntra- ve İntertümöral Heterojeniteyi Koruyan Hasta Kaynaklı Glioblastoma Organoid Modeli

Abstract

Amaç: Glioblastoma (GB), yaygın heterojenite ile karakterize edilen agresif bir beyin tümörüdür. Amacımız, histolojik özellikleri koruyan hasta kaynaklı GB organoidleri üretmek ve gelecekteki çalışmalar için bir biyobanka oluşturmaktır. Materyal ve Metot: GB hastalarından alınan tümör dokuları 0,5-1 mm'lik parçalara ayrıldı ve 14 gün boyunca organoid ortamında, orbital çalkalama altında kültüre edildi. Organoid oluşumundan önce ve sonra GFAP, OLIG2, IDH-1, p53, ATRX ve Ki67 ekspresyonunu değerlendirmek için immünohistokimyasal analiz yapıldı. Bulgular: Tümör örneklerinin tamamı 14 gün içinde küresel organoidler oluşturdu. Histolojik analiz, nükleer atipi ve pleomorfizm de dahil olmak üzere GB'ye özgü morfolojik özelliklerin korunduğunu gösterdi. GFAP, OLIG2 ve ATRX ekspresyon paternleri, ana tümörler ve organoidler arasında uyumluydu. IDH-1 ekspresyonu üç hastada tutarlıydı. Ki67 ve p53 ekspresyon seviyeleri, organoidler ve karşılık gelen tümörler arasında benzer dağılım gösterdi. Sonuç: Hasta kaynaklı GB organoidleri, temel histopatolojik özellikleri korur ve GB modellemesi için bir platform olarak hizmet edebilir.

Keywords

• Glioblastoma
• organoid kültürü
• tümör heterojenitesi

Supporting Institution

Bilimsel ve Teknolojik Araştırma Konseyi'ne (TÜBİTAK) yazar Çağla Tekin'e 2211-A doktora programı bursu kapsamında destek verdikleri için büyük teşekkür ediyoruz.

Project Number

TAY-2022-584

Ethical Statement

Bursa Uludağ Üniversitesi Etik Kurulu bu çalışmayı onaylamıştır (2021-9/14).

Thanks

İstanbul Maltepe Üniversitesi'nden Prof. Dr. Şükrü Yıldırım, Bursa Uludağ Üniversitesi'nden Prof. Dr. Şahsine Tolunay ve Doç. Dr. Mine Özsen'e IHC analizindeki destekleri için teşekkür ederiz. MRI görüntüleme konusunda özel yardımı için Prof. Dr. Bahattin Hakyemez'e de teşekkür etmek isteriz.

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