Die prognostische Relevanz von tumorinfiltrierenden Entzündungszellen im frühen Mammakarzinom

1. Hintergrund und Ziele: Die Interaktion des Immunsystems mit malignen Tumoren hat sich zu einem Schwerpunkt der onkologischen Forschung entwickelt. Große Unterschiede gibt es diesbezüglich nicht nur zwischen verschiedenen Krebsarten, sondern auch zwischen den Subtypen einer Krebsart. Dies gilt auch für das Mammakarzinom - eine tumorbiologisch äußerst heterogene Erkrankung. Das Hormonrezeptor-positive Mammakarzinom (hormone-receptor positive breast cancer, HR+BC) wurde als der am wenigsten immunogene Brustkrebs Subtyp beschrieben. Der Einfluss von tumorinfiltrierenden Entzündungszellen auf den Progress der Erkrankung sowie die Rolle der einzelnen Immunzelltypen in Bezug auf Prognose und Therapieansprechen wird daher selten untersucht und ist häufig unklar. Diese Arbeit evaluierte die Zelldichten von sechs verschiedenen Entzündungszelltypen im Tumorgewebe von Patienten mit HR+BC und setzte sie ins Verhältnis zur Langzeitprognose. Neben der grundsätzlichen Quantifizierung der Immunreaktion im Tumor stand dabei die Suche nach potenziellen prognostischen Biomarkern im Vordergrund. 2. Methoden: Das hier untersuchte Gewebe stammte von insgesamt 146 Patientinnen, deren HR+BC in den Jahren 2000 bis 2005 im Rahmen einer Phase II Studie zur beschleunigten Teilbrustbestrahlung (accelerated partial breast irradiation, APBI) am Universitätsklinikum Erlangen behandelt wurde. Stanzbiopsien aus verschiedenen Lokalisationen des Tumorgewebes mit einem Durchmesser von 2 mm wurden aus Formalin-fixierten und Paraffin-eingebetteten Operationsresektaten gewonnen und in Form von tissue micro arrays (TMAs) fixiert. Aus den TMAs wurden dann Schnitte der Dicke 4 μm produziert und mittels Immunhistochemie jeweils doppelt gefärbt, was die Markierung von zwei Typen von Immunzellen pro Schnitt erlaubte. Eine CD68/CD163 Färbung machte M1-like (CD68+ / CD163-) und M2-like (CD68+ / CD163 +) Tumor-assoziierte Makrophagen sichtbar. Eine CD4/CD45RO Färbung wurde zur Erfassung von CD4+ T-Zellen und T-Gedächtniszellen genutzt. Außerdem wurden mittels einer CD1a/CD20 Färbung unreife dendritische Zellen sowie B-Zellen im Tumor markiert. In digitalen Scans dieser gefärbten Schnitte wurden dann semiautomatisch die Flächen von Tumorepithel und Stroma sowie die Anzahl an markierten Entzündungszellen erfasst. Aus den so generierten Daten konnten Zelldichten berechnet werden, welche schließlich in Kombination mit den klinischen Daten der Patientinnen zur Überlebenszeitanalyse genutzt wurden. 3. Ergebnisse und Beobachtungen: Hohe Zelldichten von M1-like Makrophagen waren mit signifikant verbessertem krankheitsfreiem Überleben assoziiert. Für M2-like Makrophagen war das Gegenteil der Fall, hier gingen hohe Zelldichten mit einer erhöhten Rate an Fernmetastasen und Lokalrezidiven einher. Eine kombinierte Auswertung der beiden Makrophagenphänotypen ergab, dass eine Verschiebung der Makrophagenpolarisierung zum M2-like Phänotyp (M2-shifted: M1-Dichte niedrig, M2-Dichte hoch) prädiktiv für ein besonders hohes Risiko eines Lokalrezidivs bzw. einer Fernmetastasierung war. Für unreife dendritische Zellen konnte keine Korrelation mit der Prognose festgestellt werden. Hohe Zelldichten an B-Zellen und CD4+ T-Zellen waren signifikant mit reduziertem krankheitsfreiem Überleben assoziiert. Eine Infiltration des Tumors mit T-Gedächtniszellen korrelierte hingegen mit verbessertem Therapieansprechen. Ein Immunoscore basierend auf den Dichten von B-Zellen und T-Gedächtniszellen im Zentraltumor und der Invasionsfront konnte drei verschiedene Risikogruppen identifizieren. 4. Schlussfolgerungen: Trotz der gängigen Annahme, dass HR+BC als wenig immunogen gilt, war das gemessene Ausmaß der Entzündungszellinfiltration des Tumors in dieser Untersuchung teils stark mit dem krankheitsfreien Überleben assoziiert. Insbesondere die kombinierte Auswertung beider Makrophagenphänotypen in Form von Makrophagen-Shifts war ein vielversprechender potenzieller prognostischer Biomarker. Eine mögliche Erklärung für die überraschend deutlichen Ergebnisse dieser Arbeit könnte die große Homogenität der untersuchten Kohorte sein. Durch die Einschlusskriterien und Behandlungsprotokolle der APBI Studie gab es kaum Unregelmäßigkeiten bezüglich des Erkrankungsstadiums, der Tumorbiologie und der Art der Therapie. Weiterführend sind Studien in größeren Kohorten notwendig, um die hier gefundenen prognostischen Assoziationen zu bestätigen

Keine abskopalen Effekte bei Kombination von Immuntherapie und stereotaktischer Radiotherapie bei Merkel-Zell-Karzinomen

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The Prognostic Value of FoxP3+ Tumour-Infiltrating Lymphocytes in Rectal Cancer Depends on Immune Phenotypes Defined by CD8+ Cytotoxic T Cell Density

Tumour-infiltrating FoxP3+ regulatory T cells have been identified as both positive and negative prognostic factors in colorectal cancer (CRC) and rectal cancer (RC). In this study we investigated whether immune phenotypes, defined by CD8+ cytotoxic T cell density, may influence the prognostic association of FoxP3+ T cell densities in RC. Tissue microarrays from 154 rectal cancer resections were immunohistochemically double stained for CD8 and FoxP3. CD8+ and FoxP3+ cell densities were measured in the stromal and intraepithelial compartment. Stromal FoxP3+ cell densities were not associated with 10-year overall survival (OS). In the “immune-desert” phenotype, defined by very low stromal CD8+ cell density, a high density of stromal FoxP3+ T cells displayed a tendency towards an association with decreased 10-year OS (p = 0.179). In “inflamed” tumours, defined by high intraepithelial CD8+ T cell infiltration, the opposite was the case and high stromal FoxP3+ T cell densities were a positive prognostic factor (p = 0.048). Additionally, patients with an increased FoxP3/CD8 cell density ratio demonstrated a strong trend towards decreased 10-year OS (p = 0.066). These contrasting findings suggest functional heterogeneity within the group of FoxP3+ T cells. They are consistent with experimental studies which reported suppressive and non-suppressive populations of FoxP3+ T cells in CRC. Furthermore, our study demonstrates that CD8 immunohistochemistry may act as an instrument to identify tumours infiltrated by possibly functionally differing FoxP3+ T cell subtypes

Tumour-Infiltrating Inflammatory Cells in Early Breast Cancer: An Underrated Prognostic and Predictive Factor?

The role of tumour-infiltrating inflammatory cells (TIICs) in the disease progression of hormone-receptor-positive breast cancer (HR+ BC) is largely unclear since it is generally regarded as the least immunogenic BC subtype. This study investigated the prognostic significance of CD1a+ dendritic cells, CD20+ B cells, CD45RO+ memory T cells and CD4+ T-helper cells in HR+ BC. One hundred and forty-six patients were treated for early stage, distant-metastases-free HR+ BC in an accelerated partial breast irradiation (APBI) phase II trial. Immunohistochemistry was used to double-stain two adjoining sets of tissue microarrays from pre-RT (radiotherapy) tumour resection samples for CD1a/CD20 and CD45RO/CD4. Cell densities of CD1a+, CD20+, CD45RO+ and CD4+ TIICs in the stromal and intraepithelial compartment were registered semiautomatically. High densities of CD20+ and CD4+ TIICs were strongly associated with reduced disease-free survival (DFS), while high stromal CD45RO+ TIIC densities were indicators of subsequent successful treatment. An immunoscore based on CD20+ and CD45RO+ TIIC densities identified three different risk groups (p < 0.001). Thus, contrary to current assumptions, intratumoural immune cell composition might be an important prognostic indicator and a possible contributing factor in the outcome of HR+ BC and should be the subject of further research. Specifically, B-cell infiltration entailed an increased relapse rate and could play an important role in disease progression

Low-dose radiotherapy of osteoarthritis: from biological findings to clinical effects—challenges for future studies

Osteoarthritis (OA) is one of the most common and socioeconomically relevant diseases, with rising incidence and prevalence especially with regard to an ageing population in the Western world. Over the decades, the scientific perception of OA has shifted from a simple degeneration of cartilage and bone to a multifactorial disease involving various cell types and immunomodulatory factors. Despite a wide range of conventional treatment modalities available, a significant proportion of patients remain treatment refractory. Low-dose radiotherapy (LDRT) has been used for decades in the treatment of patients with inflammatory and/or degenerative diseases and has proven a viable option even in cohorts of patients with a rather poor prognosis. While its justification mainly derives from a vast body of empirical evidence, prospective randomized trials have until now failed to prove the effectiveness of LDRT. Nevertheless, over the decades, adaptions of LDRT treatment modalities have evolved using lower dosages with establishment of different treatment schedules for which definitive clinical proof is still pending. Preclinical research has revealed that the immune system is modulated by LDRT and very recently osteoimmunological mechanisms have been described. Future studies and investigations further elucidating the underlying mechanisms are an essential key to clarify the optimal patient stratification and treatment procedure, considering the patients’ inflammatory status, age, and sex. The present review aims not only to present clinical and preclinical knowledge about the mechanistic and beneficial effects of LDRT, but also to emphasize topics that will need to be addressed in future studies. Further, a concise overview of the current status of the underlying radiobiological knowledge of LDRT for clinicians is given, while seeking to stimulate further translational research.Open Access funding enabled and organized by Projekt DEAL.BMBFBMBFBMBFBayerische Forschungsstiftung http://dx.doi.org/10.13039/501100002745Universitätsklinikum Erlangen (8546

A Multicenter Evaluation of Different Chemotherapy Regimens in Older Adults With Head and Neck Squamous Cell Carcinoma Undergoing Definitive Chemoradiation

PURPOSE: The number of older adults with head-and-neck squamous cell carcinoma (HNSCC) is increasing, and treatment of these patients is challenging. Although cisplatin-based chemotherapy concomitantly with radiotherapy is considered standard regimen for patients with locoregionally advanced HNSCC, there is substantial real-world heterogeneity regarding concomitant chemotherapy in older HNSCC patients. METHODS: The XXX study is an international multicenter cohort study including older (≥65 years) HNSCC patients treated with definitive radiotherapy at 13 academic centers in the United States and Europe. Here, patients with concomitant chemoradiation were analyzed regarding overall survival (OS) and progression-free survival (PFS) using Kaplan-Meier analyses, while Fine-Gray competing risks regressions were performed regarding the incidence of locoregional failures (LRFs) and distant metastases (DMs). RESULTS: Six hundred ninety-seven patients with a median age of 71 years were included in this analysis. Single-agent cisplatin was the most common chemotherapy regimen (n=310; 44%), followed by cisplatin plus 5-fluorouracil (n=137; 20%), carboplatin (n=73; 10%), and mitomycin c plus 5-fluorouracil (n=64; 9%). Carboplatin-based regimens were associated with diminished PFS (HR=1.39 [1.03-1.89], p.05). Median cumulative dose of cisplatin was 180 mg/m2 (IQR, 120-200 mg/m2). Cumulative cisplatin doses ≥200 mg/m2 were associated with increased OS (HR=0.71 [0.53-0.95], p=.02), PFS (HR=0.66 [0.51-0.87], p=.003), and lower incidence of LRFs (SHR=0.50 [0.31-0.80], p=.004). Higher cumulative cisplatin doses remained an independent prognostic variable in the multivariate regression analysis for OS (HR=0.996 [0.993-0.999], p=.009). CONCLUSIONS: Single-agent cisplatin can be considered as the standard chemotherapy regimen for older HNSCC patients who can tolerate cisplatin. Cumulative cisplatin doses are prognostically relevant also in older HNSCC patients

Evaluation of Concomitant Systemic Treatment in Older Adults With Head and Neck Squamous Cell Carcinoma Undergoing Definitive Radiotherapy

IMPORTANCE The number of older adults with head and neck squamous cell carcinoma (HNSCC) is increasing, and these patients are underrepresented in clinical trials. It is unclear whether the addition of chemotherapy or cetuximab to radiotherapy is associated with improved survival in older adults with HNSCC. OBJECTIVE To examine whether the addition of chemotherapy or cetuximab to definitive radiotherapy is associated with improved survival in patients with locoregionally advanced (LA) HNSCC. DESIGN, SETTING, AND PARTICIPANTS The Special Care Patterns for Elderly HNSCC Patients Undergoing Radiotherapy (SENIOR) study is an international, multicenter cohort study including older adults (≥65 years) with LA-HNSCCs of the oral cavity, oropharynx/hypopharynx, or larynx treated with definitive radiotherapy, either alone or with concomitant systemic treatment, between January 2005 and December 2019 at 12 academic centers in the US and Europe. Data analysis was conducted from June 4 to August 10, 2022. INTERVENTIONS All patients underwent definitive radiotherapy alone or with concomitant systemic treatment. MAIN OUTCOMES AND MEASURES The primary outcome was overall survival. Secondary outcomes included progression-free survival and locoregional failure rate. RESULTS Among the 1044 patients (734 men [70.3%]; median [IQR] age, 73 [69-78] years) included in this study, 234 patients (22.4%) were treated with radiotherapy alone and 810 patients (77.6%) received concomitant systemic treatment with chemotherapy (677 [64.8%]) or cetuximab (133 [12.7%]). Using inverse probability weighting to attribute for selection bias, chemoradiation was associated with longer overall survival than radiotherapy alone (hazard ratio [HR], 0.61; 95% CI, 0.48-0.77; P < .001), whereas cetuximab-based bioradiotherapy was not (HR, 0.94; 95% CI, 0.70-1.27; P = .70). Progression-free survival was also longer after the addition of chemotherapy (HR, 0.65; 95% CI, 0.52-0.81; P < .001), while the locoregional failure rate was not significantly different (subhazard ratio, 0.62; 95% CI, 0.30-1.26; P = .19). The survival benefit of the chemoradiation group was present in patients up to age 80 years (65-69 years: HR, 0.52; 95% CI, 0.33-0.82; 70-79 years: HR, 0.60; 95% CI, 0.43-0.85), but was absent in patients aged 80 years or older (HR, 0.89; 95% CI, 0.56-1.41). CONCLUSIONS AND RELEVANCE In this cohort study of older adults with LA- HNSCC, chemoradiation, but not cetuximab-based bioradiotherapy, was associated with longer survival compared with radiotherapy alone

No abscopal effects of immunotherapy combined with stereotactic radiation therapy in Merkel cell carcinoma

Open Access funding enabled and organized by Projekt DEAL.Universitätsklinikum des Saarlandes und Medizinische Fakultät der Universität des Saarlandes (8981

Neutrophils in HNSCC Can Be Associated with Both a Worse or Favorable Prognosis

The prognostic significance of tumor-infiltrating neutrophils in head and neck squamous cell carcinoma (HNSCC) is poorly understood. It is unclear how the presence of neutrophils affects prognosis due to their polarization into cytotoxic N1 or immunosuppressive N2. Therefore, we determined the number of CD66b+ neutrophil granulocytes separately in the stromal and epithelial compartments in cancer tissues from 397 patients with HNSCC. Tumor samples from six historical patient groups were processed into tissue microarrays and stained immunohistochemically. In total, 21.9% were HPV positive (p16+). Neutrophil counts were much lower in the stromal compartment (372 ± 812) than in the epithelial cancer compartment (1040 ± 1477) (p < 0.001), with large differences between groups. In three groups with high neutrophil infiltration, high rates were associated with a favorable prognosis, whereas in two groups, high rates were a negative prognostic factor. In p16- oropharyngeal and hypopharyngeal cancer high infiltration was associated with a favorable prognosis. Cancers with an exclusion of neutrophils in the epithelial compartment were associated with improved prognosis. In oropharyngeal and hypopharyngeal HPV-negative cancer high neutrophil infiltration rates were clearly associated with prolonged survival. Neutrophil granulocytes in HNSCC may contribute to a favorable or unfavorable prognosis

Neutrophils in HNSCC Can Be Associated with Both a Worse or Favorable Prognosis

The prognostic significance of tumor-infiltrating neutrophils in head and neck squamous cell carcinoma (HNSCC) is poorly understood. It is unclear how the presence of neutrophils affects prognosis due to their polarization into cytotoxic N1 or immunosuppressive N2. Therefore, we determined the number of CD66b+ neutrophil granulocytes separately in the stromal and epithelial compartments in cancer tissues from 397 patients with HNSCC. Tumor samples from six historical patient groups were processed into tissue microarrays and stained immunohistochemically. In total, 21.9% were HPV positive (p16+). Neutrophil counts were much lower in the stromal compartment (372 ± 812) than in the epithelial cancer compartment (1040 ± 1477) (p < 0.001), with large differences between groups. In three groups with high neutrophil infiltration, high rates were associated with a favorable prognosis, whereas in two groups, high rates were a negative prognostic factor. In p16- oropharyngeal and hypopharyngeal cancer high infiltration was associated with a favorable prognosis. Cancers with an exclusion of neutrophils in the epithelial compartment were associated with improved prognosis. In oropharyngeal and hypopharyngeal HPV-negative cancer high neutrophil infiltration rates were clearly associated with prolonged survival. Neutrophil granulocytes in HNSCC may contribute to a favorable or unfavorable prognosis