Bu çalışma Fudan Üniversitesi Zhongshan Hastanesi'nden (2016ZSLC17) klinik araştırma özel fonu tarafından desteklendi. Yazarlar, Fudan Üniversitesi Zhongshan Hastanesi'ndeki Dr. Xianglin Hu'ya İngilizce yazmaya yönelik çok profesyonel önerileri için çok minnettarlar.

Bu tek merkezli retrospektif çalışma, Fudan Üniversitesi Zhongshan Hastanesi yerel Kurumsal İnceleme Kurulu tarafından onaylandı.
1. Bilgilendirilmiş onay alma
<ol><li>TRA müdahalelerinden önce girişimsel radyologlara (IR) TRA'nın faydalarını ve potansiyel komplikasyonlarını hastalara açıklatır.</li>
</ol>2. Hasta değerlendirmesi
<ol><li>Bilgilendirilmiş onay aldıktan sonra, ra'yı delinme ve canülasyonun fizibilitesi için değerlendirin.</li>
	<li>Hastanın tıbbi geçmişinin kapsamlı bir incelemesini gerçekleştirin. Hastaların ciddi vasküler tortuosite, şiddetli periferik vasküler hastalık, diyaliz için fistül veya RA diyaliz operasyonuna hazırlık olup olmadığını onaylayın. Bunlar TRA müdahalesi alan hastalar için göreceli kontrendikasyonlardır.</li>
	<li>RA'nın görünürlüğünü değerlendirin.<ol>
<li>Müdahaleden önce el kollateraral arterlerin ne kadar görünür olduğunu değerlendirmek için nabız oksimetresi kullanarak bir Barbeau testi gerçekleştirin11. Barbeau D dalga formu RA cannülasyonu için mutlak bir kontrendikasyon olarak kabul edilir.</li>
		<li>Barbeau C dalga formu olarak test edilen hastalar için, kol ve eldeki kol sirkülasyon miktarı hakkında daha güvenilir bilgi sağlamak için Doppler ultrason muayenesi uygulayın. 2 mm'den küçük bir iç çap kontrendikasyon olarak kabul edilir. NOT: Tıbbi öykü değerlendirmesi ve RA değerlendirmesinden sonra kontrendikasyonları olan hastalar ipsilateral RA yoluyla delinme durumundan kaçınmalıdır. Kontrallateral RA, Barbeau test değerlendirmesi yoluyla patent olduğu tespit edilirse ideal bir tedarik görevi görebilir. Sol RA başlangıçta tercih edilen bir erişim yolu olarak seçilir. Sol RA uygun bulunamazsa, sağ RA alternatif bir seçim olarak hizmet edebilir.</li>
	</ol>
</li>
</ol>3. Radyal arter erişimi
<ol><li>Hastayı anjiyografi masasına bir destek pozisyonuna getirin. Daha sonra, sol kolu hastanın vücuduna paralel ve sol bele yakın bir yere yerleştirin, kateter ve telin kolay yerleştirilmesini sağlar ve operatör konumlandırmasını TFA ile karşılaştırılabilir hale getirin.</li>
	<li>Distal RA nabzını palpasyonla işaretleyin. Cilt yüzeyini %10 povidon-iyot cerrahi ovma çözeltisi ile temizleyin ve çözeltinin havayla kurumasını bekleyin. Sol kolu cerrahi bir örtü ile örtün.<ol>
<li>Olası sol RA delinme arızası durumunda, sağ kol veya sağ kasık bölgesini sterilize ederek ve perdeleyerek alternatif bir erişim yolu hazırlayın.</li>
	</ol>
</li>
	<li>Lokal anestezi uygulayın (yani% 2 1 mL lidoksin) sol RA'nın en güçlü titreşim ekseni boyunca styloid işlemine proksimal.</li>
	<li>Modifiye Seldinger tekniğini kullanarak bileği uzatın ve RA'yı 20 G iğne ile delin. Pulsatile arteriyel kan dönüşü göze çarpıldığında, 0.025 inç hidrofilik kılavuz tel tanıtın.<ol>
<li>Kılavuz telini geri çek ve dirençle karşılaşılırsa iğneyi yeniden düzenle. Kılavuz telin takılmasını zorlamayın. Dijital çıkarma anjiyografisinin (DSA) yardımıyla, RA'yı vurgulamak ve hidrofilik kılavuz telini yerleştirmeye yardımcı olmak için yaklaşık 1 ila 2 mL kontrast enjekte edin.</li>
	</ol>
</li>
	<li>Erişim elde edildikten sonra, iğneyi çıkarın ve kılavuz tel ile 4 Fransız hidrofilik kılıfı tanıtın. Kılış takıldıktan sonra, kılım ucunun damar içinde olduğunu doğrulamak için bir şırınna ile az miktarda arteriyel kanı hafifçe geri pompalayın.</li>
</ol>4. Antikoagülasyon ve vasküler genişleme
<ol><li>10 mL vazodilasyon kokteyl çözeltisi hazırlayın (3.000 IU kırılmamış heparin, 0.1 mg nitrogliserin ve 20 mg lidotrin).</li>
	<li>Vazodilasyon kokteyl çözeltisinin 8 mL'lik kısmını kılıktan 0,5 mL/s hızında geçirin(Şekil 1). NOT: Orta veya yüksek kanama riski olan hastalar için heparin dozunu azaltın veya durdurun.</li>
</ol>5. Kateter seçimi
<ol><li>Subklaviyen arterden geçmek ve alçalanı meşgul etmek için 4-Fransız, 125 cm ortak kateter ve standart 0,035 inç x 180 cm hidrofilik tel kullanın. Bir arter döngüsüne veya damar işkencesine olası lezyonları önlemek için kol içinde navigasyon sırasında proksimal aksiller arteri görselleştirmek için DSA floroskopi kullanın. NOT: Subklaviyen arterin birçok atardamar dalı vardır. Anjiyografik kılavuz, retrograd kateterizasyon sırasında kılavuz tel kateterlerin kollateral damarlara girmesini önler. Birkaç vakada radyal arterde atardamar döngüsü vardır. Standart tel döngüyü geçemiyorsa, bir mikrokreter kullanılması ve açılı 0.016 inç veya 0.018 inç mikro tel kullanılması önerilir.</li>
	<li>4-Fransız, 125 cm ortak kateteri standart bir 0.035 inç tel ile birlikte kullanarak kılavuz telleri alçalana doğru yönlendirmek için enine kemeri müzakere edin. NOT: Aort ve sol subklaviyen arter arasındaki açı çok akutsa, bu dönüşü gerçekleştirmek için Kobra 2 şeklinde bir kateter (örneğin Simmons I veya Simmons II kateter) önerilir.</li>
	<li>İnen aortu kateterizasyondan sonra, ortak kateteri koaksiyel bir teknikle değiştirin. Ortak kateter alçalan aort içine yerleştirildikten sonra, DSA floroskopisi rehberliğinde çölyak gövdesinin kateterizasyonu için kateter ucunu ventral olarak yönlendirin. Çoğu durumda, çölyak gövdesinin, hepatik arterin ve üstün mezenterik arterin kateterize ve anjiyografisini yapmak kolaydır. NOT: Çölyak arteri ile azalan aort arasındaki açı çok akutsa, işlemi tamamlamak için bir Kobra kateteri kullanın.</li>
	<li>Hepatik embolizasyon işlemleri için koaksiyel bir teknik kullanarak süper seçici kateterizasyon ve kemoembolizasyon gerçekleştirin ve tümörleri besleyen hepatik arterin hedeflenen dalına 2,8-Fransız 150 cm mikrokreter yerleştirin (Şekil 2). Hastalığın yüküne ve hasta tercihine göre TACE işlemi gerçekleştirin.</li>
	<li>Yeterli embolizasyonu doğrulamak için yüksek basınçlı bir enjektör kullanarak ortak kateter aracılığıyla anjiyogram yapın. Kateter ucu genellikle ortak hepatik arterde bulunur. Kontrast maddesinin 9-12 mL'lik kısmını 3-4 mL oranında, floroskopi süresi ~15 s olan enjekte edin. Ardından, RA'nın zarar görmesini önlemek için kateteri bir kılavuz telin üzerine çıkarın.</li>
</ol>6. Radyal arter hemostazı
NOT: Nonocclusive hemostaz RA açıklığını korumak için özel bir turnike kullanılarak gerçekleştirilir (Şekil 3).
<ol><li>Kalan 2 mL vazodilasyon kokteyl çözeltisini (bölüm 4) RA kılımı aracılığıyla geçirin. Hemen sonra, kılım yaklaşık 5 cm alın.</li>
	<li>Sol bilekteki radyal erişim bölgesinin üzerine bir turnike yerleştirin ve beraberindeki şırınnayı kullanarak turnike hava yastığını hava ile yeterince şişirin. Sonra kılıfi tamamen çıkarın ve hava yastığını yavaşça söndür. Erişim alanında sızıntı gözlendiğinde, manşete 1 mL hava ekleyin. Tipik olarak, hemostaz tutmak için hava yastığına 10-15 mL hava eklenir.</li>
	<li>Kanama veya sızıntı olmadığını onaylayın. Aynı zamanda, distal radyal arter darbesinin hemostaz sırasında hissedilebilir olduğundan emin olun. Sol başparmak üzerindeki arteriyel dalga biçimini doğrulamak için darbe oksimetre dalga biçimini kullanın.</li>
	<li>Hava kesesini her 2 saatte bir ~2 mL'de 6 saatten uzun süre yavaşça şişirin. Ameliyattan 6 saat sonra turnike çıkarıldıktan sonra hemostazın gerçekleştirildiğini doğrulayın. NOT: Deflasyon sırasında delinme bölgesinden kanama veya sızıntı görülürse, hava kesesine 30 dakika boyunca hava eklenir ve işlem tekrarlanır.</li>
	<li>Taburcu olmadan önce, RA'nın açıklığını doğrulamak ve radyal arter tıkanıklığı olan hastaları kaydetmek ve yakından takip etmek için Barbeau testi yaptırın.</li>
</ol>7. Takip
<ol><li>Müdahaleden yaklaşık 1 ay sonra, TRA hastalarına sol bilek muayenesi ve nabız muayenesi de dahil olmak üzere kapsamlı bir fizik muayene verin. Potansiyel olarak tıkanmış RA'ları olan hastalar için, forearm Doppler ultrason veya nabız oksimetresi kullanarak el kan akışının sonraki değerlendirmelerini yapın.</li>
	<li>TACE sonrası tüm hastaları yakından takip edin. BT taramalarında yeni tümör nodülleri belirginse ve ilk lezyonlar yeniden yaygınlaşmış gibi görünüyorsa, başka bir TACE tedavisi yapın.</li>
</ol>

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L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transradial+Approach+for+Hepatic+Radioembolization:+Initial+Results+and+Technique.">Transradial Approach for Hepatic Radioembolization: Initial Results and Technique.</a> AJR: American Journal of Roentgenology. 207 (5), 1112-1121 (2016).</li><li>Bernat, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ST-segment+elevation+myocardial+infarction+treated+by+radial+or+femoral+approach+in+a+multicenter+randomized+clinical+trial:+the+STEMI-RADIAL+trial.">ST-segment elevation myocardial infarction treated by radial or femoral approach in a multicenter randomized clinical trial: the STEMI-RADIAL trial.</a> Journal of the American College of Cardiology. 63 (10), 964-972 (2014).</li></ol>

WZ ve ZPY tarafından çalışma konsepti ve tasarımı; ND, ZHZ ve MJY tarafından veri alımı; ZPY tarafından fon elde edildi. Yazarların konuyla ilgili herhangi bir finansal açıklaması bulunmamaktadır.

TRA girişimsel tedavi son yıllarda özellikle tanısal ve Girişimsel kardiyoloji işlemlerinde dünya çapında önemli ölçüde artmıştır12. Ayrıca periferik vasküler hastalık müdahalesine de ilgi artmaktadır. Prosedürel başarı oranlarından ödün vermeden, TRA'dan kardiyak girişime TFA13 , 14ile karşılaştırıldığında kanama ve vasküler komplikasyon oranlarını etkili bir şekilde azaltabilir. TFA ile karşılaş...

Hepatosellüler karsinom (HCC), dünya çapında altıncı en yüksek insidans oranına sahip çok yaygın bir malignitedir. Aynı zamanda dünyada kanser ölümlerinin ikinci önde gelen nedenidir1. Hastaların sadece %5-20'si iyileştirici tedavi alabildiğinden, transarterial kemoembolizasyon (TACE) tedavi edilemeyen HCC2hastaları için en popüler palyatif tedavidir. TACE,3.ara evrede HCC hastaları için en sık kullanılan ve etkili tedavi yaklaşımı olarak kabul edilmiştir. Transfemoral erişim (TFA) kemoembolizasyonu TACE4için en yaygın yaklaşımdır. Bununla birlikte, erişim yerinde kanama ve majör vasküler komplikasyonlar da dahil olmak üzere TFA müdahalesi ile ilgili riskler vardır5. Bu komplikasyonlar hastaneye yatış süresinin uzamasına ve maliyetlerin artmasına yol açar. Ayrıca, TFA en az 6 saat boyunca hareketsiz hale getirilmesini gerektirir, bu da hastalar için rahatsızlık ve memnuniyetsizliği arttırır.
Transradial erişim (TRA), yirmi yıldan fazla bir süredir perkütan koroner müdahalede (PCI) kullanılan alternatif bir yaklaşımdır5,6. TRA PCI'nin birkaç avantajı vardır: artan prosedür konforu, erişim bölgesine bağlı kanamada azalma, majör vasküler komplikasyonlarda azalma ve mortalitede azalma7,8. Radyal artere (RA) yüzeysel konumu nedeniyle erişmek ve delinmek kolaydır7. Hemostazın müdahaleden sonra yapılması kolaydır ve katı bir immoblizasyon yoktur9. Kardiyak kateterizasyonda TRA müdahalesi için teşvik edici kanıtlara rağmen, bugüne kadar sadece birkaç çalışmada TRA periferik hastalık müdahalesinde kullanılmıştır. Kötü huylu karaciğer tümörleri için TRA müdahaleleri daha da nadirdir. Burada TRA hepatik embolizasyonun klinik fizibilitesi ve güvenliği analiz edilir. Bir kurumun sağlanan adım adım TRA protokolü ile ilgili deneyimi de açıklanmıştır.

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			<td height="21" style="height:21px;width:141px;">Pharmorubicin</td>
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			<td height="42" style="height:42px;width:141px;">Maestro Microcatheter</td>
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			<td height="21" style="height:21px;width:141px;">Sheath Introducer</td>
			<td style="width:127px;">Merit</td>
			<td style="width:123px;">PSI-4F-11-035</td>
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			<td height="21" style="height:21px;width:141px;">Steerable Guidewire</td>
			<td style="width:127px;">Merit</td>
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			<td height="21" style="height:21px;width:141px;">TR Band</td>
			<td style="width:127px;">Terumo</td>
			<td style="width:123px;">XX*RF06</td>
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			<td style="width:127px;">SonoScape</td>
			<td style="width:123px;">20172231180</td>
			<td></td>
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transradial access chemoembolization for hepatocellular carcinoma patients

Transarterial kemoembolizasyon (TACE), hepatosellüler karsinom (HCC) tedavisinde ara aşamada en sık görülen modalitedir. TACE genellikle transfemoral erişim (TFA) yoluyla gerçekleştirilir. Ancak koroner arter müdahalelerinde komplikasyon ve mortalitenin azalmasına bağlı olarak transradyal erişim (TRA) tercih edilmektedir. TRA'nın avantajlarının TACE gerekli araştırmaya uygulanıp uygulanamayacağı.
Tek merkezde TRA TACE alan hastalar retrospektif olarak çalışmaya alındı. Prosedürel detaylar, teknik başarı, radyal arter tıkanma (RAO) oranı ve erişim bölgesine bağlı kanama komplikasyonları değerlendirildi. Ekim 2017'den Ekim 2018'e kadar 112 hastaya 160 TRA TACE işlemi uygulandı. Genel teknik başarı oranı %95,0 (152/160) idi. TRA'dan TFA'ya geçiş oranı %1,9 oldu. Hiçbir vakada erişim bölgesine bağlı kanama komplikasyonlarına rastlanmadı. Asemptomatik RA tıkanma üç hastada (%2.7) meydana geldi. TFA ile karşılaştırıldığında TRA, erişim sitesine bağlı kanama komplikasyonlarını azaltırken güvenliği ve hasta memnuniyetini artırabilir. Ayrıca, TRA müdahaleleri ileri yaş, obezite veya kanama komplikasyonları riski yüksek olan hastalara fayda sağlayabilir.

Ekim 2017'den Ekim 2018'e kadar 112 hastaya 160 TRA TACE işlemi uygulandı ve genel teknik başarı oranı %95,0 (152/160) oldu. Sekiz vaka teknik arızayla karşılandı. Bunlardan beşi sol RA delinme hatasından kaynaklandı ve daha sonra sağ RA erişimi ile başarılı TACE uygulandı. Diğer üç vaka kanülasyon hatasından kaynaklandı ve daha sonra sağ SK erişimine çapraz geçişle başarılı bir müdahaleye maruz kaldı. RA erişiminin SK erişimine geçiş oranı sadece% 1,9 idi. Vakaların hiçbirinde er...

Watch this Scientific Journal Video about Transradial Access Chemoembolization for Hepatocellular Carcinoma Patients at JoVE.com

Transradial Access Chemoembolization for Hepatocellular Carcinoma Patients

Transarterial kemoembolizasyon (TACE), hepatosellüler karsinomun ara aşamasındaki hastalar için standart tedavidir ve tipik olarak femoral arter erişimi ile gerçekleştirilir. Transfemoral erişim ile karşılaştırıldığında, transradial erişim (TRA) kanama komplikasyonlarının oranını azaltabilir ve hasta toleransını artırabilir. Burada radyal arter erişimi ile transarterial kemoembolizasyon yapmak için bir yöntem sunulmaktadır.

Hepatosellüler Karsinom Hastaları için Transradyal Erişim Kemoembolizasyonu

Transarterial chemoembolization (TACE) is the most common modality for treatment of hepatocellular carcinoma (HCC) at the intermediate stage. TACE is ...

transradial-access-chemoembolization-for-hepatocellular-carcinoma

Research

JoVE Journal

Cancer Research

5.3K Görüntüleme.  Fudan University. Transarterial kemoembolizasyon (TACE), hepatosellüler karsinomun ara aşamasındaki hastalar için standart tedavidir ve tipik olarak femoral arter erişimi ile gerçekleştirilir. Transfemoral erişim ile karşılaştırıldığında, transradial erişim (TRA) kanama komplikasyonlarının oranını azaltabilir ve hasta toleransını artırabilir. Burada radyal arter erişimi ile transarterial kemoembolizasyon yapmak için bir yöntem sunulmaktadır.

Video: Transradial Access Chemoembolization for Hepatocellular Carcinoma Patients

Study of Viral Vectors in a Three-dimensional Liver Model Repopulated with the Human Hepatocellular Carcinoma Cell Line HepG2

This protocol describes the generation of a three-dimensional (3D) ex vivo liver model and its application to the study and development of viral vector systems. The model is obtained by repopulating the extracellular matrix of a decellularized rat liver with a human hepatocyte cell line. The model permits studies in a vascularized 3D cell system, replacing potentially harmful experiments with living animals. Another advantage is the humanized nature of the model, which is closer to human physiology than animal models.
In this study, we demonstrate the transduction of this liver model with a viral vector derived from adeno-associated viruses (AAV vector). The perfusion circuit that supplies the 3D liver model with media provides an easy means to apply the vector. The system permits monitoring of the major metabolic parameters of the liver. For final analysis, tissue samples can be taken to determine the extent of recellularization by histological techniques. Distribution of the virus vector and expression of the delivered transgene can be analyzed by quantitative PCR (qPCR), Western blotting and immunohistochemistry. Numerous applications of the vector model in basic research and in the development of gene therapeutic applications can be envisioned, including the development of novel antiviral therapeutics, cancer research, and the study of viral vectors and their potential side effects.

The recellularized extracellular matrix of a decellularized rat liver can be used as a humanized, three-dimensional ex vivo model to study the distribution and transgene expression of a virus or viral vector.

Insan hepatoselüler karsinoma hücre hattı HepG2 ile üç boyutlu Karaciğer modeli yeniden doldurulur Viral Vektörler Study

This protocol describes the generation of a three-dimensional (3D) ex vivo liver model and its application to the study and development of viral vector ...

Kanser Araştırmaları

A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma

Perineural invasion (PNI) is found in approximately 40% of head and neck squamous cell carcinomas (HNSCC). Despite multimodal treatment with surgery, radiation, and chemotherapy, locoregional recurrences and distant metastases occur at higher rates, and overall survival is decreased by 40% compared to HNSCC without PNI. In vitro studies of the pathways involved in HNSCC PNI have historically been challenging given the lack of a consistent, reproducible assay. Described here is the adaptation of the dorsal root ganglion (DRG) assay for the examination of PNI in HNSCC. In this model, DRG are harvested from the spinal column of a sacrificed nude mouse and placed within a semisolid matrix. Over the subsequent days, neurites are generated and grow in a radial pattern from the cell bodies of the DRG. HNSCC cell lines are then placed peripherally around the matrix and invade preferentially along the neurites toward the DRG. This method allows for rapid evaluation of multiple treatment conditions, with very high assay success rates and reproducibility.

Perineural invasion (PNI) is a common feature of head and neck squamous cell carcinoma (HNSCC), conferring lower survival rates. Its mechanisms are poorly understood. Utilizing neurites generated from murine dorsal root ganglia confined to a semisolid matrix, the pathways involved in the PNI of HNSCC cell lines can be investigated.

Baş ve Boyun Skuamöz Hücre Karsinomu içinde perinöral invazyon İçin Bir Model

Perineural invasion (PNI) is found in approximately 40% of head and neck squamous cell carcinomas (HNSCC). Despite multimodal treatment with surgery, ...

Automated Multiplex Immunofluorescence Panel for Immuno-oncology Studies on Formalin-fixed Carcinoma Tissue Specimens

Continued developments in immuno-oncology require an increased understanding of the mechanisms of cancer immunology. The immunoprofiling analysis of tissue samples from formalin-fixed, paraffin-embedded (FFPE) biopsies has become a key tool for understanding the complexity of tumor immunology and discovering novel predictive biomarkers for cancer immunotherapy. Immunoprofiling analysis of tissues requires the evaluation of combined markers, including inflammatory cell subpopulations and immune checkpoints, in the tumor microenvironment. The advent of novel multiplex immunohistochemical methods allows for a more efficient multiparametric analysis of single tissue sections than does standard monoplex immunohistochemistry (IHC). One commercially available multiplex immunofluorescence (IF) method is based on tyramide-signal amplification and, combined with multispectral microscopic analysis, allows for a better signal separation of diverse markers in tissue. This methodology is compatible with the use of unconjugated primary antibodies that have been optimized for standard IHC on FFPE tissue samples. Herein we describe in detail an automated protocol that allows multiplex IF labeling of carcinoma tissue samples with a six-marker multiplex antibody panel comprising PD-L1, PD-1, CD68, CD8, Ki-67, and AE1/AE3 cytokeratins with 4&#8242;,6-diamidino-2-phenylindole as a nuclear cell counterstain. The multiplex panel protocol is optimized in an automated IHC stainer for a staining time that is shorter than that of the manual protocol and can be directly applied and adapted by any laboratory investigator for immuno-oncology studies on human FFPE tissue samples. Also described are several controls and tools, including a drop-control method for fine quality control of a new multiplex IF panel, that are useful for the optimization and validation of the technique.

A detailed protocol for a six-marker multiplex immunofluorescence panel is optimized and performed, using an automated stainer for more consistent results and a shorter procedure time. This approach can be directly adapted by any laboratory for immuno-oncology studies.

IMMUNO-Onkoloji araştırmalar Formalin sabit Karsinomu doku örnekleri Multiplex ayirt Panel otomatik

Continued developments in immuno-oncology require an increased understanding of the mechanisms of cancer immunology. The immunoprofiling analysis of ...

Multidimensional Coculture System to Model Lung Squamous Carcinoma Progression

Tumor-stroma interactions play a critical role in the development of lung squamous carcinoma (LUSC). However, understanding how these dynamic interactions contribute to tissue architectural changes observed during tumorigenesis remains challenging due to the lack of appropriate models. In this protocol, we describe the generation of a 3D coculture model using a LUSC primary cell culture known as TUM622. TUM622 cells were established from a LUSC patient-derived xenograft (PDX) and have the unique property to form acinar-like structures when seeded in a basement membrane matrix. We demonstrate that TUM622 acini in 3D coculture recapitulate key features of tissue architecture during LUSC progression as well as the dynamic interactions between LUSC cells and components of the tumor microenvironment (TME), including the extracellular matrix (ECM) and cancer-associated fibroblasts (CAFs). We further adapt our principal 3D culturing protocol to demonstrate how this system could be utilized for various downstream analyses. Overall, this organoid model creates a biologically rich and adaptable platform that enables one to gain insight into the cell-intrinsic and extrinsic mechanisms that promote the disruption of epithelial architectures during carcinoma progression and will aid the search for new therapeutic targets and diagnostic markers.

An in vitro model system was developed to capture tissue architectural changes during lung squamous carcinoma (LUSC) progression in a 3-dimensional (3D) co-culture with cancer-associated fibroblasts (CAFs). This organoid system provides a unique platform to investigate the roles of diverse tumor cell-intrinsic and extrinsic changes that modulate the tumor phenotype.

Akciğer Skuamöz Karsinom Progresyonunu Modellemek Için Çok Boyutlu Coculture Sistemi

Tumor-stroma interactions play a critical role in the development of lung squamous carcinoma (LUSC). However, understanding how these dynamic interactions ...

Analysis of Liver Microenvironment During Early Progression of Non-Alcoholic Fatty Liver Disease-Associated Hepatocellular Carcinoma in Zebrafish

Liver cancer is currently the third&#160;leading cause of cancer related death worldwide, and Hepatocellular Carcinoma (HCC) accounts for 75-90% of all liver cancer cases. With the introduction of effective treatments to prevent and treat hepatitis B/C, non-alcoholic fatty liver disease (NAFLD), and the more aggressive form know as non-alcoholic steatohepatitis (NASH), are quickly becoming the number one risk factors to develop HCC in modern societies. To better understand the role NASH has on the development of HCC we designed a NASH-associated HCC zebrafish. The optical clarity and genetic tractability of the zebrafish larvae make them an appealing and powerful model to study the liver microenvironment and immune cell composition using non-invasive fluorescent live imaging. This protocol describes how to use a NASH-associated HCC zebrafish model to investigate the effect of cholesterol surplus in the liver microenvironment and its impact on immune cell composition at early stages of the disease. First, we feed HCC larvae (s704Tg), which express hepatocyte-specific activated beta-catenin, with a 10% high cholesterol diet for 8 days to develop a NASH-associated HCC model. Here we describe how to make use of different transgenic lines to evaluate several early malignancy features in the liver by non-invasive confocal microscopy, such as liver area, cell, and nuclear morphology (hepatocytes area, nuclear area, nuclear:cytoplasmic ratio (N:C ratio), nuclear circularity, micronuclei/nuclear herniation scoring) and angiogenesis. Then, using transgenic lines with tagged immune cells (neutrophils, macrophages, and T cells) we show how to analyze liver immune cell composition in NASH-associated HCC larvae. The described techniques are useful to evaluate liver microenvironment and immune cell composition at early hepatocarcinogenesis stages, but they can also be modified to study such features in other liver disease models.

Here, we present how to generate a non-alcoholic fatty liver disease (NAFLD)-associated Hepatocellular Carcinoma (HCC) zebrafish model to study the impact of cholesterol surplus on liver microenvironment and immune cell landscape.

Zebra Balığında Alkolsüz Yağlı Karaciğer Hastalığına Bağlı Hepatosellüler Karsinomun Erken İlerlemesi Sırasında Karaciğer Mikroçevresinin Analizi

Liver cancer is currently the third&#160;leading cause of cancer related death worldwide, and Hepatocellular Carcinoma (HCC) accounts for 75-90% of all ...

A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma

The aggressiveness and lack of well-tolerated and widely effective treatments for advanced hepatocellular carcinoma (HCC), the predominant form of liver cancer, rationalize its rank as the second most common cause of cancer-related death. Preclinical models need to be adapted to recapitulate the human conditions to select the best therapeutic candidates for clinical development and aid the delivery of personalized medicine. Three-dimensional (3D) cellular spheroid models show promise as an emerging in vitro alternative to two-dimensional (2D) monolayer cultures. Here, we describe a 3D tumor spheroid&#160;model which exploits the ability of individual cells to aggregate when maintained in hanging droplets, and is more representative of an in vivo environment than standard monolayers. Furthermore, 3D spheroids can be produced by combining homotypic or heterotypic cells, more reflective of the cellular heterogeneity in vivo, potentially enabling the study of environmental interactions that can influence progression and treatment responses. The current research optimized the cell density to form 3D homotypic and heterotypic tumor spheroids by immobilizing cell suspensions on the lids of standard 10 cm3 Petri dishes. Longitudinal analysis was performed to generate growth curves for homotypic versus heterotypic tumor/fibroblasts spheroids. Finally, the proliferative impact of fibroblasts (COS7 cells) and liver myofibroblasts (LX2) on homotypic tumor (Hep3B) spheroids was investigated. A seeding density of 3,000 cells (in 20 &#181;L media) successfully yielded Huh7/COS7 heterotypic spheroids, which displayed a steady increase in size up to culture day 8, followed by growth retardation. This finding was corroborated using Hep3B homotypic spheroids cultured in LX2 (human hepatic stellate cell line) conditioned medium (CM). LX2 CM triggered the proliferation of Hep3B spheroids compared to control tumor spheroids. In conclusion, this protocol has shown that 3D tumor spheroids can be used as a simple, economical, and prescreen in vitro tool to study tumor-stromal interactions more comprehensively.

Comprehensive in vitro models that faithfully recapitulate the relevant human disease are lacking. The current study presents three-dimensional (3D) tumor spheroid creation and culture, a reliable in vitro tool to study the tumor-stromal interaction in human hepatocellular carcinoma.

Hepatosellüler Karsinomdaki Tümör-Stromal Etkileşimi Araştırmak İçin Üç Boyutlu Sferoid Modeli

The aggressiveness and lack of well-tolerated and widely effective treatments for advanced hepatocellular carcinoma (HCC), the predominant form of liver ...

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

Hepatosellüler Karsinomun Ortotopik Fare Modelinde Non-İnvaziv PET/MR Görüntüleme

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

Establishment and Characterization of Patient-Derived Xenograft Models of Anaplastic Thyroid Carcinoma and Head and Neck Squamous Cell Carcinoma

Patient-derived xenograft (PDX) models faithfully preserve the histological and genetic characteristics of the primary tumor and maintain its heterogeneity. Pharmacodynamic results based on PDX models are highly correlated with clinical practice. Anaplastic thyroid carcinoma (ATC) is the most malignant subtype of thyroid cancer, with strong invasiveness, poor prognosis, and limited treatment. Although the incidence rate of ATC accounts for only 2%-5% of thyroid cancer, its mortality rate is as high as 15%-50%. Head and neck squamous cell carcinoma (HNSCC) is one of the most common head and neck malignancies, with over 600,000 new cases worldwide each year. Herein, detailed protocols are presented to establish PDX models of ATC and HNSCC. In this work, the key factors influencing the success rate of model construction were analyzed, and the histopathological features were compared between the PDX model and the primary tumor. Furthermore, the clinical relevance of the model was validated by evaluating the in vivo therapeutic efficacy of representative clinically used drugs in the successfully constructed PDX models.

The present protocol establishes and characterizes a patient-derived xenograft (PDX) model of anaplastic thyroid carcinoma (ATC) and head and neck squamous cell carcinoma (HNSCC), as PDX models are rapidly becoming the standard in the field of translational oncology.

Anaplastik Tiroid Karsinomu ve Baş-Boyun Skuamöz Hücreli Karsinomun Hasta Kaynaklı Ksenogreft Modellerinin Oluşturulması ve Karakterizasyonu

Patient-derived xenograft (PDX) models faithfully preserve the histological and genetic characteristics of the primary tumor and maintain its ...

Hedef belirleme ve ilaç geliştirme için HCC organoid oluşumunun optimize edilmesi

Development and Optimization of A Human Hepatocellular Carcinoma Patient-Derived Organoid Model for Potential Target Identification and Drug Discovery

Hepatocellular carcinoma (HCC) is a highly prevalent and lethal tumor worldwide and its late discovery and lack of effective specific therapeutic agents necessitate further research into its pathogenesis and treatment. Organoids, a novel model that closely resembles native tumor tissue and can be cultured in vitro, have garnered significant interest in recent years, with numerous reports on the development of organoid models for liver cancer. In this study, we have successfully optimized the procedure and established a culture protocol that enables the formation of larger-sized HCC organoids with stable passaging and culture conditions. We have comprehensively outlined each step of the procedure, covering the entire process of HCC tissue dissociation, organoid plating, culture, passaging, cryopreservation, and resuscitation, and provided detailed precautions in this paper. These organoids exhibit genetic similarity to the original HCC tissues and can be utilized for diverse applications, including the identification of potential therapeutic targets for tumors and subsequent drug development.

Yazar Spotlight: Hasta Kaynaklı Organoidler Kullanarak Karaciğer Kanseri Patogenezinin Araştırılması 

We provide a comprehensive overview and refinement of existing protocols for hepatocellular carcinoma (HCC) organoid formation, encompassing all stages of organoid cultivation. This system serves as a valuable model for the identification of potential therapeutic targets and the assessment of drug candidate effectiveness.

Potansiyel Hedef Belirleme ve İlaç Keşfi için İnsan Hepatoselüler Karsinomu Hasta Kaynaklı Organoid Modelinin Geliştirilmesi ve Optimizasyonu

Hepatocellular carcinoma (HCC) is a highly prevalent and lethal tumor worldwide and its late discovery and lack of effective specific therapeutic agents ...