Title Prognostička vrijednost izražaja antigena za rak testisa i tumorskog mikrookoliša u trostruko negativnim karcinomima dojke Title (english) Prognostic value of testicular cancer antigen expression and tumor microenvironment in triple-negative breast cancers Author Toni Čeprnja Mentor Ivana Mrklić (mentor)Committee member Ivana Kuzmić Prusac (predsjednik povjerenstva)Committee member Branka Petrić Miše (član povjerenstva)Committee member Katarina Vukojević (član povjerenstva)Granter University of Split Defense date and country 2023-06-28, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 616 - Pathology. Clinical medicine Abstract Cilj istraživanja. Ciljevi ovog istraživanja bili su utvrditi povezanost povećanog broja
stromalnih limfocita (sTIL), intratumoralnih limfocita (itTIL), limfatičnih nakupina i
izražaja antigena za rak testisa (CTA) u trostruko negativnim karcinomima dojke s
dužinom preživljenja bez znakova povrata bolesti (DFS). Također, željeli smo istražiti
utjecaj izražaja CTA na broj TIL, formiranje limfatičnih nakupina i pojavu „basal-like“
morfoloških karakteristika u ovoj skupini karcinomima dojke. Dodatno smo željeli
utvrditi povezanost analiziranih pokazatelja s kliničko-patološkim prognostičkim
pokazateljima kao što su dob, klinički stadij, veličina tumora, histološki gradus,
histološki podtip, „basal-like“ morfologija i proliferacijska aktivnost mjerena
primjenom Ki-67 protutijela.
Materijali i metode. U studiju je uključeno 97 bolesnica s dijagnozom ranog TNBC-a
koje su u razdoblju u razdoblju od 1. siječnja 2017. do 31. prosinca 2018. godine
operirane u četiri Klinička bolnička centra, kod kojih nije provedeno neoadjuvantno
liječenje. Na hemalaun-eozinom obojenim patohistološkim uzorcima analizirana su
obilježja tumora i tumorskog mikrookoliša. Uzorci su dodatno bojani
imunohistokemijski primjenom protutijela za PD-L1 (SP-142) i CTA (NY-ESO-1,
MAGE A1 i multi MAGE A). Pri očitavanju izražaja CTA korištene su dvije granične
vrijednosti: granična vrijednost određena prema medijanu određenog CTA u ispitivanoj
skupini, te arbitrarno postavljena granična vrijednost od ≥10% ukupnog broja tumorskih
stanica.
Za usporedbu kategorijskih varijabli korišten je χ2 test i test logističke regresije. Za
usporedbu kvantitativnih varijabli i ispitivanih karakteristika tumora korišten je Mann-
Whitney test. Coxovom regresijskom analizom utvrđena je povezanost istraživanih
varijabli s dužinom DFS bolesnika (u mjesecima). U analizi preživljenja korištena je
Kaplan-Meierova krivulja preživljenja i log-rank test.
Rezultati. PD-L1 ICS ≥1% zabilježen je u 47 uzoraka (49%), a statistički je značajno
korelirao s histološkom gradusom, povećanim brojem sTIL i formiranjem LN. BL
morfološke karakteristike zabilježene su u 47 (49%) TNBC, a statistički su značajno
korelirale s višim Ki-67 proliferacijskim indeksom (p=0.001) i višim histološkim
gradusom (p=0.005). Na osnovu visoke razine sTIL 11 TNBC (11%) svrstano je u LP
skupinu, a bili su statistički značajno povezani s pozitivnim PD-L1 ICS (p=0.001),
višim Ki-67 proliferacijskim indeksom (p=0.04) te većim brojem itTIL (p<0.001).
Izražaj NY-ESO-1 bio je pozitivan u 37 TNBC (38%), uz značajnu korelaciju s
povećanim brojem itTIL (p = 0.039). Izražaj multi MAGE A bio je pozitivan u 77
(79%) TNBC, a korelirao je s pojavom BL morfologije. Dulje preživljenje bez znakova
povrata bolesti (DFS) bilo je statistički značajno povezano s negativnim limfnim
čvorovima (p=0.011) te prisutnošću primarnih LN (p = 0.027). Iako nismo uspjeli
dokazati statističku značajnost, niti jedna od ispitanica s intratumoralnim TIL ≥10%
(N=12), te s prisutnoću sekundarnih limfatičnih nakupina (N=14) nije imala znakove
povrata bolesti u intervalu praćenja od 48 mjeseci.
Zaključak. Dokazali smo da prisutnost primarnih LN ima prognostički značaj u
trostruko negativnim karcinomima dojke, te da identifikacija i karakterizacija imunih
stanica u tumorskom mikrookolišu na osnovu smještaja, stupnja organizacije i odnosa s
tumorskim stanicama može pomoći u dodatnoj subklasifikaciji trostruko negativnih
tumora u različite prognostičke skupine.
CTA se smatraju potencijalnim kandidatima za ciljano liječenje i za razvoj
antitumorskih cjepiva. Tome u prilog govori i pozitivan izražaj NY-ESO-1 zabilježen u
38% TNBC u ovom istraživanju, uz statistički značajnu povezanost s povećanim brojem
itTIL
Razumijevanje biologije trostruko negativnih tumora i njegove interakcije s
mikrookolišem, uz korelaciju sa markerima kao što su PD-L1 i CTA, koji mogu kočiti
ili poticati imuni odgovor, donosi nova saznanja koja mogu doprinjeti unaprijeđenju
načina liječenja za ovu terapijski vrlo ograničenu skupinu tumora.
Abstract (english) Objectives. The aim of this study was to determine the association of increased number
of stromal lymphocytes (sTIL), intratumoral lymphocytes (itTIL), lymphoid aggregates
and expression of cancer testis antigen (CTA) in triple negative breast cancers with
disease-free survival (DFS). We also wanted to investigate the influence of CTA
expression on the amount of TIL, the formation of lymphoid aggregates and the
occurance of "basal-like" morphological characteristics in this group of breast cancers.
Additionally, we wanted to study the relationship of the analyzed variables with
clinical-pathological prognostic characteristic such as age, clinical stage, tumor size,
histological grade, histological subtype, "basal-like" morphology and Ki-67
proliferation activity.
Materials and methods. The study included 97 patients with a diagnosis of early
TNBC who underwent surgery in four Clinical Hospital Centers between January 1st,
2017 and December 31st, 2018, with no neoadjuvant treatment administration. Tumor
characteristics and tumor microenvironment were analyzed on hemalaun-eosin stained
pathohistological slides. The samples were additionally immunostained using antibodies
for PD-L1 (SP-142) and CTA (NY-ESO-1, MAGE A1 and multi MAGE A). In
evaluating CTA expression, two cut-off values were used: a value determined according
to the median value for each CTA in the examined group, and an arbitrarily set value of
≥10%.
The χ2 test and the logistic regression test were used to compare categorical variables.
The Mann-Whitney test was used to compare quantitative variables and examined
tumor characteristics. Cox regression analysis was used to determine the association of
all investigated variables with the DFS (in months). For the survival analysis, the
Kaplan-Meier survival curve and the log-rank test were used.
Results. PD-L1 ICS ≥1% was recorded in 47 samples (49%), and statistically
significantly correlated with histological grade, increased number of sTIL and LA
formation. BL morphological characteristics were recorded in 47 (49%) TNBC, and
statistically significantly correlated with higher Ki-67 proliferation index (p=0.001) and
higher histological grade (p=0.005). Based on the level of sTIL, 11 TNBCs (11%) were
assigned to the LP group and were statistically significantly associated with positive
PD-L1 ICS (p=0.001), higher Ki-67 proliferation index (p=0.04) and higher number of
itTIL (p<0.001). NY-ESO-1 expression was positive in 37 TNBCs (38%), with a
significant correlation with increased number of itTIL (p = 0.039). The expression of
multi MAGE A was positive in 77 (79%) TNBC and correlated with occurance of BL
morphology. Longer disease-free survival (DFS) was associated with negative lymph
nodes (p=0.011) and the presence of primary LA (p = 0.027). Although we failed to
prove statistical significance, none of the patients with intratumoral TIL ≥10% (N=12)
and with the presence of secondary LA (N=14) had signs of disease recurrence in 48
months long follow-up period.
Conclusion. We proved that the presence of primary LA has a prognostic significance
in the triple negative breast cancer, so identification and characterization of immune
cells in the tumor microenvironment based on location, degree of organization and
relationship with tumor cells can help in additional subclassification of triple-negative
cancers into different prognostic groups.
CTAs are considered potential candidates for targeted treatment and for the
development of antitumor vaccines. This is supported by the positive expression of NYESO-
1 recorded in 38% of TNBC in this study, with a statistically significant
association with increased number of itTIL.
Understanding of triple-negative breast cancer biology and its interaction with the
microenvironment, along with correlations with markers such as PD-L1 and CTA,
which can inhibit or stimulate the immune response, brings new knowledge that can
contribute to the improvement of treatment for this group of tumors with limited
treatment options.
Keywords
Trostruko negativne novotvorine dojke
Novotvorine testisa
Tumorsko mikrookruženje
Keywords (english)
Triple Negative Breast Neoplasms
Testicular Neoplasms
Tumor Microenvironment
Language croatian URN:NBN urn:nbn:hr:171:838248 Study programme Title: Biology of Neoplasms Type of resource Text File origin Born digital Access conditions Open access Terms of use Created on 2023-06-13 07:27:51
