Microsatellite unstable (MSI) colorectal cancer (CRC) tumors have a high mutational load (particularly frame-shift mutations) that creates numerous neoantigens that are presented to major histocompatibility complex molecules and recognized by T cells. Consequently, MSI tumors have a higher presence of tumor-infiltrating lymphocytes than mismatch repair-proficient tumors. Colorectal cancer patients with MSI constitute a rare group of immune checkpoint inhibitor (ICI)-responsive patients. Nonetheless, complete radiological responders comprise between 3% and 16% of MSI advanced CRC patients, which compares poorly with the 45% to 87% rate of pathological complete response in early MSI CRC patients treated with ICIs. In this review, we address the efficacy of current ICIs and the biological differences between early and advanced MSI CRC to potentially increase the efficacy of ICIs in both settings.

Background: Adjuvant radiotherapy and hormonotherapy after breast-conserving surgery (BCS) in ductal carcinoma in situ (DCIS) have been shown to reduce the risk of local recurrence. To predict the risk of ipsilateral breast tumor relapse (IBTR) after BCS, the Memorial Sloan Kettering Cancer Center (MSKCC) developed a nomogram to analyze local recurrence (LR) risk in our cohort and to assess its external validation. Methods: A historical cohort study using data from 296 patients treated for DCIS at the Hospital Clínic of Barcelona was carried out. Patients who had had a mastectomy were excluded from the analysis. Results: The mean age was 58 years (42–75), and the median follow-up time was 10.64 years. The overall local relapse rate was 13.04% (27 patients) during the study period. Actuarial 5- and 10-year IBTR rates were 5.8 and 12.9%, respectively. The external validation of the MSKCC nomogram was performed using a multivariate logistic regression analysis on a total of 207 patients, which did not reach statistical significance in the studied population for predicting LR (p = 0.10). The expression of estrogen receptors was significantly associated with a decreased risk of LR (OR: 0.25; p = 0.004). Conclusions: In our series, the LR rate was 13.4%, which was in accordance with the published series. The MSKCC nomogram did not accurately predict the IBTR in this Spanish cohort of patients treated for DCIS (p = 0.10).

INTRODUCTION: Breast cancer (BC) is a highly prevalent and heterogeneous disease, entailing different so-called intrinsic subtypes (IS) according to gene expression, namely Luminal A, Luminal B, HER2-Enriched (HER2E) and Basal-like, as well as a normal breast-like group. The HER2E is still a poorly understood entity, which needs further biologic characterization to improve therapeutic management. MATERIAL AND METHODS: Patients (pts) treated at Hospital Clinic (Barcelona, Spain) over 18 years with a diagnosis of metastatic BC, with available matched primary and metastatic tumor samples for gene expression analyses were retrospectively recruited. Using the nCounter® Breast Cancer 360 panel, we studied the expression of 776 genes pertaining to different tumor and immune pathway-related signatures, with a focus on HER2E vs. non-HER2E tumors. Significant changes were considered at a false discovery rate (FDR) < 5%. Main clinicopathological features were also compared. IS changes from primary to metastatic disease were assessed. RESULTS: Ninety-one pts with paired tumor samples were included. Briefly, primary tumors were 67.0% hormone receptor-positive (HR+)/HER2-negative (HER2-), 14.8% were HER2-positive (HER2+) and 18.2% were triple-negative (TNBC). IS distribution in primary tumors was the following: 25 Luminal A (28%), 22 Luminal B (24%), 24 HER2E (26%), 12 Basal-like (13%), and 8 Normal-like (9%). In contrast, IS distribution in metastatic tumors was the following: 13 Luminal A (14%), 22 Luminal B (24%), 34 HER2-E (37%), 16 Basal-like (18%), and 6 Normal-like (7%). The HER2E disease proved to be a relatively stable subtype, with 16 (66.7%) tumors not changing IS in the transition to metastatic disease (p=0.078). Particularly, within HR+/HER2-, HER2+ and TNBC, 8/12 (66.7%), 8/10 (80.0%) and 1/2 (50.0%) tumors remained HER2E. Conversely, an overall significant switch from Luminal to non-Luminal tumors at the metastatic progression was observed (p=0.031), with 14/19 (73.7%) new non-Luminal BC being HER2E. When considering all primary and metastatic tumors, HER2E were observed to be less frequently estrogen receptor (ER) positive than non-HER2E tumors (55.8% vs. 78.7%; p=0.002), with lower mean progesterone receptor (PR) levels (15.3% vs. 25.8%; p=0.027). Compared to the other subtypes (as a group), the PAM50 risk of recurrence score (ROR-P) was higher for HER2E tumors (59.1 vs. 41.7; p < 0.001), which were also more frequently HER2+ (36.5% vs. 5.8%) and less likely HR+/HER2- (50.0% vs. 73.6%) and TNBC (13.5% vs. 20.7%), compared to non-HER2E (p < 0.001). No significant differences in grade, TILs, Ki67 and histotype were observed. Overall, 140/776 genes were significantly downregulated in HER2E vs. non-HER2E tumors, including genes related to cell adhesion, migration, DNA damage repair, apoptosis, estrogen signalling pathway, senescence. Conversely, 178/776 genes were significantly upregulated, including the tyrosine-kinase receptor FGFR4, genes involved in nuclear activity, DNA transcription regulation, MAP-kinase signaling pathways, proliferation, cytokine response, epithelial-to-mesenchymal transition (EMT), cell cycle progression, and immune-related genes such as CD274 (PD-L1 gene). DISCUSSION: A switch towards more aggressive IS from primary to advanced disease was observed, with an increase in HER2E prevalence. HER2E tumors, which tend to maintain their IS at the metastatic disease, showed upregulation of genes related to proliferation, survival and EMT, coherently with their well-known worse prognosis. FGFR4 and CD274 upregulation, along with downregulation of genes involved in DNA damage repair suggest these tumors might benefit from targeted therapeutic approaches. Genomic higher risk was not convoyed by consistent clinicopathological features, except for lower PR levels and HER2 overexpression at IHC. Further characterization according to primary/metastatic and HR status is ongoing. Intrinsic subtype distribution across primary and metastatic breast tumors Citation Format: Francisco Javier Muñoz-Carrillo, Laia Paré, Benedetta Conte, Adela Rodríguez, Patricia Galván, Esther Sanfeliu, Blanca González-Farré, Claudette Falato, Isabel Garcia-Fructuoso, Barbara Adamo, Nuria Chic, Olga Martínez-Sáez, Tomás Pascual, Reinaldo Moreno, Montserrat Muñoz, Fara Brasó-Maristany, Aleix Prat, Francesco Schettini, Maria Vidal. Clinico-Pathological and Molecular Characterization of HER2-Enriched Breast Tumors Independently of HER2 Status [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-23-16.

Background and purpose Oxaliplatin-induced neuropathy (OIN) implies axonal damage of both small and large sensory nerve fibers. We aimed at comparing the neurophysiological changes occurred after treatment and the capability to recovery based on histological marker of re-innervation GAP-43. Methods 48 patients with cancer were assessed before and after chemotherapy (at 3 months and 12 months if available). We recorded ulnar and sural sensory nerve action potentials (SNAP), determined quantitative sensory thresholds for warm and cold (WDT, CDT), pain thresholds and collected a distal biopsy of skin to assess the intra-epidermal nerve fiber density (IENFD) with PGP9.5 and GAP-43 markers (in a subgroup of 19 patients). Results Increased WDT and CDT as well as diminished IENFD at distal leg were already found in 30% of oncologic patients before treatment. After oxaliplatin, there was a significant increase in thermal thresholds in 52% of patients, and a decrease of SNAP amplitude in the sural nerve in 67% patients. IENFD was reduced in 47% and remained unchanged in 37% after oxiplatin. The density of GAP-43 + fibers and GAP-43/PGP 9.5 ratio was similar before and after treatment showing that cutaneous re-innervation is preserved despite no clinical recovery was observed after one year. Conclusion Non-selective axonal loss affects sensory fibers in OIN. However, the presence of intra-epidermal regenerative sprouts detected by GAP-43 may reduce the impact of neurotoxicity in the small fibers with long-term sequelae mostly on myelinated nerve endings. Pre-oxaliplatin GAP-43 failed to identify patients with higher risk of damage or worse recovery after treatment.

In advanced breast cancer, biomarker identification and patient selection using a metastatic tumor biopsy is becoming more necessary. However, the biology of metastasis according to the organ site is largely unknown. Here, we evaluated the expression of 771 genes in 184 metastatic samples across 11 organs, including liver, lung, brain, and bone, and made the following observations. First, all PAM50 molecular intrinsic subtypes were represented across organs and within immunohistochemistry-based groups. Second, HER2-low disease was identified across all organ sites, including bone, and HER2 expression significantly correlated with ERBB2 expression. Third, the majority of expression variation was explained by intrinsic subtype and not organ of metastasis. Fourth, subtypes and individual subtype-related genes/signatures were significantly associated with overall survival. Fifth, we identified 74 genes whose expression was organ-specific and subtype-independent. Finally, immune profiles were found more expressed in lung compared to brain or liver metastasis. Our results suggest that relevant tumor biology can be captured in metastatic tissues across a variety of organ sites; however, unique biological features according to organ site were also identified and future studies should explore their implications in diagnostic and therapeutic interventions.

Next–generation sequencing (NGS) provides a molecular rationale to inform prognostic stratification and to guide personalized treatment in cancer patients. Here, we determined the prognostic and predictive value of actionable mutated genes in metastatic colorectal cancer (mCRC). Among a total of 294 mCRC tumors examined by targeted NGS, 200 of them derived from patients treated with first–line chemotherapy plus/minus monoclonal antibodies were included in prognostic analyses. Discriminative performance was assessed by time–dependent estimates of the area under the curve (AUC). The most recurrently mutated genes were TP53 (64%), KRAS or NRAS (49%), PIK3CA (15%), SMAD4 (14%), BRAF (13%), and FBXW7 (9.5%). Mutations in FBXW7 correlated with worse OS rates (p = 0.036; HR, 2.24) independently of clinical factors. Concurrent mutations in TP53 and FBXW7 were associated with increased risk of death (p = 0.02; HR, 3.31) as well as double–mutated TP53 and SMAD4 (p = 0.03; HR, 2.91). Analysis of the MSK–IMPACT mCRC cohort (N = 1095 patients) confirmed the same prognostic trend for the previously identified mutated genes. Addition of the mutational status of these genes upon clinical factors resulted in a time–dependent AUC of 87%. Gene set enrichment analysis revealed specific molecular pathways associated with SMAD4 and FBXW7 mutations in TP53–defficient tumors. Conclusively, SMAD4 and FBXW7 mutations in TP53–altered tumors were predictive of a negative prognostic outcome in mCRC patients treated with first–line regimens.

Abstract Background: There is an increasing need in the clinic to biopsy metastatic disease in patients with advanced breast cancer (ABC). However, the microenvironment and tumor cell biology of breast cancer metastasis is largely unknown. Here, we report a molecular characterization of ABC according to the site of metastasis. Methods: RNA from 184 FFPE metastatic samples were evaluated using the nCounter BC 360 Panel, which includes the expression of 689 BC-related genes and 82 immune-related genes. PAM50 subtypes (Basal-like, HER2-enriched [HER2-E], Luminal A, Luminal B and Normal-like) were also determined. HER2 protein expression was assessed by immunohistochemistry (IHC) in 115 tumor samples, and HER2-low tumors (i.e. 1+ or 2+ and ISH-negative) were identified. Descriptive statistics, significance analysis of microarrays (using False Discovery Rate [FDR]) and logistic regressions were used to identify organ-specific gene expression profiles. Finally, we derived an organ-specific predictor of 209-genes from our cohort, and applied it to the RNAseq-based TCGA PanCancer dataset, which includes 174 glioblastomas multiforme, 424 liver hepatocellular carcinomas and 576 lung adenocarcinomas, among other cancer-types. Results: A single metastatic tumor sample from 184 individual patients with ABC was obtained from bone (18%), liver (17%), skin (15%), brain (12%), breast (13%), lymph nodes (9%), lung (7%), pleura (5%), ovary (2%), muscle (1%) and peritoneum (1%). All PAM50 subtypes were identified across the main organ sites; however, significant differences in subtype distribution were observed (p<0.001). Basal-like subtype was more prevalent in brain, lung and skin metastasis (FDR=0.3%). Unsupervised analysis and principal component analysis revealed brain and liver metastasis as the most distinct. Supervised analysis identified organ-specific genes independently of PAM50 subtype, i.e.: bone-specific genes (WIF1, IBSP, MMP9 and ITGB3); brain-specific genes (CRYAB, SOX10, FGF1 and CHI3L1); liver-specific genes (ALDH1A1, CYP4F3, PCK1, and SFRP2); lung-specific genes (CAV1, WNT5A, PTGS2 and IL6); skin-specific genes (KRT14, KRT5, S100A7 and SERPINB5). Among the organ-specific gene list, 15 (30%) of the 50 PAM50 genes were identified, including up-regulation of FGFR4 in liver, ESR1 in bone, ERBB2 in lung and KRT5 and KRT14 in skin. Regarding ERBB2, a high correlation between ERBB2 mRNA and HER2 IHC expression (0, 1+, 2+ and 3+) was observed (p<0.001). Interestingly, HER2-low disease was identified across all PAM50 subtypes and organ sites, including bone metastasis. Regarding immune-genes, all were found differentially expressed across the main organ sites (FDR<5%) with lung metastasis showing the highest expression (i.e. PDCD1, CD8A, GMZA, IL1B) and liver and brain metastasis the lowest. Finally, the 209-gene organ-specific predictor in PanCancer TCGA identified 96% of glioblastomas multiforme as brain, 98.6% of liver hepatocellular carcinomas as liver and 57.1% of lung adenocarcinoma as lung. Conclusions: The main sites of metastasis in ABC have unique biological features independently of tumor molecular subtype. Our results suggest that treatment strategies based on the site(s) of metastasis should be explored. Citation Format: Fara Brasó-Maristany, Laia Paré, Núria Chic, Olga Martínez-Sáez, Tomás Pascual, Meritxell Mallafré, Blanca González-Farré, Esther Sanfeliu, Débora Martínez, Patricia Galván, Belinda Salinas, Barbara Adamo, Reinaldo Moreno, Maria Vidal, Montserrat Muñoz, Aleix Prat. Molecular characterization of advanced breast cancer according to site of metastasis [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS18-11.

Circulating tumor DNA (ctDNA) levels may predict response to anticancer drugs, including CDK4/6 inhibitors and endocrine therapy combinations (CDK4/6i+ET); however, critical questions remain unanswered such as which assay or statistical method to use. Here, we obtained paired plasma samples at baseline and week 4 in 45 consecutive patients with advanced breast cancer treated with CDK4/6i+ET. ctDNA was detected in 96% of cases using the 74-gene Guardant360 assay. A variant allele fraction ratio (VAFR) was calculated for each of the 79 detected mutations between both timepoints. Mean of all VAFRs (mVAFR) was computed for each patient. In our dataset, mVAFR was significantly associated with progression-free survival (PFS). Baseline VAF, on-treatment VAF or absolute changes in VAF were not associated with PFS, nor were CA-15.3 levels at baseline, week 4 or the CA-15.3 ratio. These findings demonstrate that ctDNA dynamics using a standardized multi-gene panel and a unique methodological approach predicts treatment outcome. Clinical trials in patients with an unfavorable ctDNA response are needed.

We read with interest the study of Monti et al ,1 the first rheumatic disease cases with COVID-19. In detail, the authors described the clinical course of COVID-19 in a series of 11 patients with rheumatoid arthritis, one with psoriatic arthritis and one with spondyloarthritis treated with immunosuppressive targeted therapies. Here, we describe the main characteristics of four patients with Behcet’s disease (BD) with COVID-19. Data on patients with systemic autoimmune diseases with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection are currently lacking. Data from the first 110 patients included in the COVID-19 Global Rheumatology Alliance and the European League Against Rheumatism (EULAR)–COVID-19 Database have been recently published.2 Here we describe, to our knowledge, the first single-centre experience of COVID-19 in patients who fulfilled the international criteria for BD,3 including clinical characteristics, antiviral and immunomodulatory treatment, and outcomes. All patients gave informed consent for publishing their clinical data. We used nasopharyngeal swab samples for all diagnoses, amplifying …

3553 Background: Several prognostic clinical scores for metastatic colorectal cancer (mCRC) GEMCAD (Ann Oncol 25 (Supp 4)), 2014; GERCOR (Oncologist 16, 2011); Köhne (Ann Oncol 13, 2002) are useful for treatment guidance. Next generation sequencing (NGS) allows the evaluation of multiple deregulated pathways (WNT, TGFB, PI3K-RTK-RAS and p53 signaling) (Nature, 2012). We hypothesize that complex vs simple genotypes evaluated by NGS, could be distributed differently according sidedness and clinical scores and would provide independent prognostic value.Methods: NGS by Ion Torrent in 22 CRC significant genes was employed to process samples from 141 consecutive mCRC patients (pts) diagnoses from February 2016 to November 2017 in a single institution. Genotype was defined as Complex:1) BRAF mutant 2) RTK-RAS-PI3K+/-p53+FBXW7 mutant 3) RTK-RAS-PI3K+/-p53+SMAD4 mutant; Simple: 1) RTK-RAS-PI3K mutant alone 2) p53 mutant alone 3) RTK-RAS-PI3K mutant+p53 mutant 4) no mutations. Associations were analyzed with Fisher-t test. Cox proportional hazard models and interaction analyses were used to explored the effect of genotype, primary sidedness and clinical scores with overall survival (OS). Results: Informative cases (128/141); 91%. Complex genotype; 29%; p53 (67%), KRAS (45%), BRAF (14%), SMAD4 (13%), PI3K (12%), FBXW7 (8%), NRAS (3%), other ( < 3%). Complex genotype was not associated with sidedness (p = 0.997) but was associated with high-risk GEMCAD score (p = 0.03). In the Cox model, genotypes by NGS remain significant for OS independently of clinical scores (see Table).Conclusions: NGS genotype provides independent prognostic information beyond clinical scores. The combination of both variables should allow optimal prognostic stratification in mCRC. Prognostic value for overall survival (cox proportional hazard model). GEMCAD GERCOR Köhne HR (95% IC) p HR (95% IC) p HR (95% IC) p Genotype Simple 1 1 1 Complex 1.82 (1.04 – 3.20) 0.037 2.13 (1.22 – 3.73) 0.008 2.31 (1.31 – 4.08) 0.004 Score Low 1 1 1 Intermediate 2.96 (0.67 – 12.94) 0.149 5.32 (0.72 – 39.39) 0.101 2.13 (0.94 – 4.78) 0.067 High 7.83 (1.84 – 33.34) 0.005 14.81 (2.01 –109.40) 0.008 7.78 (3.69 – 16.40) 0.0001