Translational Oncology 38 (2023) 101782

Contents lists available at ScienceDirect

Translational Oncology
journal homepage: www.elsevier.com/locate/tranon

Original Research

Correlation of an immune-related 8-gene panel with pathologic response to

neoadjuvant chemotherapy in patients with primary breast cancers
Ling-Ming Tseng a, b, c, d, Chi-Cheng Huang a, c, e, Yi-Fang Tsai a, b, c, Ji-Lin Chen a,
Ta-Chung Chao a, b, f, Jiun-I Lai f, g, h, Pei-Ju Lien a, i, Yen-Shu Lin a, c, Chin-Jung Feng a,
Yen-Jen Chen a, b, c, Jen-Hwey Chiu a, c, j, Chih-Yi Hsu b, k, Chun-Yu Liu a, b, f, l, *
a
Comprehensive Breast Health Center, Taipei Veterans General Hospital, Taipei, Taiwan
b
School of Medicine, National Yang Ming Chiao Tung University, Hsinchu, Taiwan
c
Division of General Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
d
Division of Experimental Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
e
Department of Public Health, College of Public Health, National Taiwan University, Taipei, Taiwan
f
Division of Medical Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan
g
Center of Immuno-Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan
h
Institute of Clinical Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan
i
Department of Nursing, Taipei Veterans General Hospital, Taipei, Taiwan
j
Institute of Traditional Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan
k
Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital, Taipei, Taiwan
l
Division of Transfusion Medicine, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan

A R T I C L E I N F O A B S T R A C T

Keywords: Neoadjuvant chemotherapy (NACT)-induced pathologic complete response (pCR) is associated with a favorable
Neoadjuvant chemotherapy prognosis for breast cancer. Prior research links tumor-infiltrating lymphocytes with breast cancer chemotherapy
Breast cancer response, suggesting the tumor-immune microenvironment’s role. The aim of this study was to evaluate the
Pathologic complete response
immune-related genes that exhibit associations with the response to NACT. In this study, we analyzed a total of
37 patients (aged 27–67) who received NACT as the first-line treatment for primary breast cancer, followed by
surgery. This group consisted of nine patients (24.3 %) with estrogen receptor (ER)-positive/HER2-negative
status, ten patients (27.0 %) with ER-positive/HER2-positive status, five patients (13.5 %) with ER-negative/
HER2-positive status, and thirteen patients (35.1 %) with triple-negative breast cancer (TNBC). Among these
patients, twelve (32.4 %) achieved a pCR, with eight (66.6 %) having HER2-positive tumors, and the remaining
four having TNBC. To identify immune-related genes linked with pCR in subjects with breast cancer prior to
NACT, we collected fresh tissues for next-generation sequencing. Patients with pCR had higher expressions of
eight genes, KLRK1, IGJ, CD69, CD40LG, MS4A1, CD1C, KLRB1, and CA4, compared to non-pCR patients. The 8-
gene signature was associated with good prognosis and linked to better relapse-free survival in patients receiving
chemotherapy. The expression of these genes was involved in better drug response, displaying a positive cor­
relation with the infiltration of immune cells. In conclusion, we have identified eight immune-related genes that
are associated with a favorable prognosis and positive responses to drugs. This 8-gene signature could potentially
provide prognostic insights for breast cancer patients undergoing NACT.

Introduction adjuvant treatments (administered after breast surgery) and neo­

adjuvant treatments (given before surgery). Combinations of surgery,
In breast cancer treatment, the local approach involves surgery fol­ chemotherapy, endocrine therapy, and targeted therapy have the po­
lowed by post-operative radiation. Systemic therapy encompasses tential to cure early-stage breast cancer. However, advanced-stage

* Corresponding author at: Division of Transfusion Medicine, Department of Medicine, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei
112, Taiwan.
E-mail address: cyliu3@vghtpe.gov.tw (C.-Y. Liu).

https://doi.org/10.1016/j.tranon.2023.101782
Received 27 June 2023; Received in revised form 23 August 2023; Accepted 6 September 2023
Available online 13 September 2023
1936-5233/© 2023 Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
L.-M. Tseng et al. Translational Oncology 38 (2023) 101782

Table 1 long-term disease-free survival, particularly in HER2-positive breast

Association of pCR with clinicopathological characteristics in breast cancer cancer and TNBC [10,11]. The commercially available Oncotype DX
patients. Breast Recurrence Score test is a genomic-based comprehensive assess­
Characteristic non-pCR (n = 25) pCR (n = 12) P value ment for patients with ER-positive, HER2-negative tumors in the NACT
Age 0.232
setting [12]. However, the reliable biomarkers for evaluating pCR re­
≤60 18 (72.0) 11 (91.7) mains limited.
>60 7 (28.0) 1 (8.3) The significant role of the immune system in both conventional
AJCC TNM stage 0.384 chemotherapy and targeted therapy has been reported [13]. Numerous
I 1 (4.0) 0 (0.0)
studies and clinical trials have explored the changes in the tumor
II 6 (24.0) 4 (33.3)
III 18 (72.0) 7 (58.3) microenvironment during NACT for breast cancer. Notably, a high level
IV 0 (0.0) 1 (8.3) of tumor-infiltrating lymphocytes (TILs) has been identified as an in­
Tumor stage (T) 0.033 dependent prognostic marker for achieving pCR [14,15]. Moreover,
T1 3 (12.0) 1 (8.3) there exists a positive correlation between pCR and higher expression of
T2 7 (28.0) 9 (75.0)
T3 6 (24.0) 2 (16.7)
immune-related genes at baseline prior to NACT. High expression of
T4 9 (36.0) 0 (0.0) chemoattractant cytokines and cytotoxic T cell markers has been asso­
Nodal status (N) 0.707 ciated with achieving pCR [16]. Patients with a pre-existing or induced
N0 6 (24.0) 1 (8.3) immune response show a better prognosis than who lack either trait
N1 6 (24.0) 4 (33.3)
[17]. However, the relationship between tumor-immune interactions
N2 7 (28.0) 4 (33.3)
N3 6 (24.0) 3 (25.0) and inflammatory genes concerning NACT-induced pCR in breast cancer
ER 0.495 remains ambiguous. To bridge this gap, we employed targeted RNA
Positive 14 (56.0) 5 (41.7) sequencing, focusing on the identification of immune-related genes
Negative 11 (44.0) 7 (58.3) associated with pCR. This endeavor aims to facilitate the advancement
PR 0.084
of targeted therapies and predictive biomarkers. We examined the gene
Positive 12 (48.0) 2 (16.7)
Negative 13 (52.0) 10 (83.3) expressions of breast cancer tissues prior to NACT using a
HER2 0.036 next-generation sequencing-based immuno-oncology platform, with the
Positive 7 (28.0) 8 (66.7) goal of uncovering factors connected to pCR. We ascertained the base­
Negative 18 (72.0) 4 (33.3)
line expression of genes involved in tumor-immune interactions and
Subtypes 0.058
ERþ/HER2- 9 (36.0) 0 (0.0) examined their correlation with outcome.
ERþ/HER2þ 5 (20.0) 5 (41.7)
ER-/HER2þ 2 (8.0) 3 (25.0) Methods
TNBC 9 (36.0) 4 (33.3)
Histologic grade 0.542
Patients and specimens
1 2 (8.0) 0 (0.0)
2 17 (68.0) 8 (66.7)
3 6 (24.0) 4 (33.3) This VGH-TAYLOR study (NCT04626440) was approved and granted
by the ethics committee of the Institutional Review Board of Taipei
Veterans General Hospital, Taipei City, Taiwan (No. 2018-09-007A). All
breast cancer, including triple-negative breast cancer (TNBC), is asso­
participants provided written informed consent to take part in this
ciated with a less favorable prognosis. Neoadjuvant chemotherapy
study, which was conducted in compliance with the Helsinki Declara­
(NACT) has gained widespread acceptance as a strategy to shrink tumors
tion. Participants eligible for this study include females aged 20 years or
and target micrometastases in locally advanced breast cancer. This
older with a confirmed diagnosis of primary invasive breast cancer, who
approach aims to reduce the extent of disease before surgery [1,2].
are planning to undergo breast cancer treatments. This study also con­
NACT-induced pathologic complete response (pCR) has been associated
siders subjects with breast cancer recurrence at screening or stage IV
with favorable prognosis for patients with breast cancer. Response to
subjects currently undergoing or having received breast cancer treat­
NACT is associated with prolonged disease-free survival and possibly
ments. Participants should have an Eastern Cooperative Oncology Group
overall survival. As a result, pCR has been employed to evaluate treat­
performance status of 3 or lower, a life expectancy of at least 3 months,
ment response and has functioned as a surrogate endpoint in clinical
and provide written informed consent. However, those with a primary
trials [3,4]. Although there has been some uncertainty with respect to
cancer other than breast cancer within the past 5 years will be excluded
the surrogacy between pCR and long-term survival [5,6], regulatory
from participation [18]. From December 2018 to October 2020, a total
agencies have adopted pCR as a surrogate endpoint for long-term out­
of 37 female patients who received NACT as the first-line treatment for
comes in early-stage breast cancer [7]. DNA and germline status have
breast cancer were recruited. The study included 12 subjects with pCR
been found to be associated with the response to NACT. For instance, the
and 25 subjects who not achieved a pathologic complete response
GeparSixto trial illustrates that the inclusion of neoadjuvant carboplatin
(non-pCR) or had breast cancer recurrence. TNM staging, histological
alongside a regimen comprising anthracycline, taxane, and bev­
grade, and tumor subtypes were determined in accordance with the
acizumab leads to increased rates of pCR in patients with TNBC.
WHO classification system. Clinical data were collected from medical
Importantly, within this context, patients possessing germline mutations
records by oncologists. A summary of patient demographics was pre­
in BRCA1 and BRCA2 showed improved pCR rates in the absence of
sented in Table S1.
carboplatin, whereas patients lacking BRCA1 and BRCA2 mutations
exhibited better pCR rates in the presence of carboplatin [8].
RNA sequencing
Clinically, breast cancer subtyping is generally based on the protein
expression levels of estrogen receptor (ER), progesterone receptor (PR),
The Oncomine Immune Response Research Assay (Thermo Fisher
and human epidermal growth factor receptor 2 (HER2). Given the het­
Scientific, Waltham, MA, USA) was utilized to determine tumor-immune
erogeneity of breast cancer, different subtypes exhibit distinct clinical
interactions. Fresh tumor tissues were subjected to total RNA extraction,
phenotypes and behaviors [9]. Consequently, the search for novel ap­
and 10 ng of this RNA was reverse transcribed into cDNA using the
proaches to improve the rate of pCR and to identify breast cancer pa­
SuperScript VILO cDNA Synthesis Kit (Thermo Fisher Scientific). Li­
tients who could benefit from NACT remains a challenge. Meta-analyses
braries were prepared using the Ion AmpliSeq Library Kit 2.0 and the
have shown that pCR serves as a robust biomarker for predicting
Oncomine Immune Response Research Assay (Thermo Fisher Scientific)

2
L.-M. Tseng et al.
Fig. 1. The association of the 8-gene signature with response to NACT. (A) Volcano plots of differential gene expression in the VGH-TAYLOR study. Each dot
represents the average value of a gene, the x-axis is the Log2-transformed fold change for the non-pCR to pCR ratio, whereas the y-axis shows Log10-transformed P-
value. (B) A Heatmap represents the DEGs between patients with pCR and non-pCR. (C and D) Receiver operating characteristic curve of the eight DEGs and 8-gene
signature with the AUC in the VGH-TAYLOR study (C) and GSE20194 cohort (D).
following the manufacturer’s instructions. Template preparation and
sequencing were performed using the Ion Chef system and Ion S5
sequencing system. Gene expression levels were quantified using the
Torrent Suite immuneResponseRNA plugin. The analysis encompassed
fold changes of 395 genes distributed across 36 functional annotation
groups, including lymphocyte regulation, cytokine signaling, check­
point pathways, and tumor characterization [19].
In silico analyses with public databases
For validation, an independent cohort (GSE20194) comprising 278
breast cancer patients who received NACT, including paclitaxel, 5-fluo­
rouracil, cyclophosphamide, and doxorubicin, was subjected to analysis
[20]. Relapse-free survival (RFS) data for breast cancer patients who
underwent chemotherapy were examined from the Kaplan-Meier plotter
and the Molecular Taxonomy of Breast Cancer International Consortium
(METABRIC) databases [21,22]. RFS curves were plotted against time,
based on transcript levels. The correlation between gene expression and
the response to NACT in breast cancer patients was analyzed using the
ROC plotter [23]. Drug sensitivity and gene expression data of cancer
cell lines in the Cancer Therapeutics Response Portal were consulted.
Additionally, pathway analysis was conducted using GSCALite [24,25].
Gene transcript levels of breast cancer tissues and normal tissues from
The Cancer Genome Atlas (TCGA) were downloaded and analyzed [26].
The correlation between gene expression and the infiltration level of
immune cells was investigated using the TIMER database [27].
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Translational Oncology 38 (2023) 101782
Statistical analysis
The mean of gene expression in non-pCR and pCR groups was
calculated using Tukey’s bi-weight average algorithm. For survival
analysis, RFS curves of breast cancer patients receiving chemotherapy
were plotted using the Kaplan-Meier method and compared using the
Log-rank test. The RFS events included the first recurrence of invasive
disease (at a local, regional, or distant site) and death from any cause.
Patients who did not experience RFS events were treated as censored at
their most recent follow-up. Cox proportional hazards regression anal­
ysis was employed to evaluate the influence of immune-related genes on
RFS. Comparisons between non-pCR and pCR groups were performed
using contingency tables, and the chi-square test was employed for
clinicopathological characteristics. Logistic regression was utilized to
ascertain the predictive significance of clinicopathological characteris­
tics for pCR. All statistical analyses were performed using SPSS for
Windows software, version 22.0 (SPSS, Chicago, IL, USA). A P-value <
0.05 was considered statistically significant.
Results
Eight immune-related gene expressions are higher in breast cancer patients
with pCR
To explore the association between immune-related gene expressions
and pCR in breast cancer patients receiving NACT, a total of 37 patients
were enrolled. The median age of these patients was 51 years, ranging
from 27 to 67 years. This cohort comprised nine patients (24.3 %) with
ER-positive/HER2-negative tumors, ten patients (27.0 %) with ER-
L.-M. Tseng et al.
Fig. 2. The 8-gene signature is associated with good prognosis in breast cancer. The Kaplan–Meier analysis of the expressions of eight DEGs and 8-gene on RFS in
patients with breast cancer receiving chemotherapy from the KM plotter database.
positive/HER2-positive tumors, five patients (13.5 %) with ER-
negative/HER2-positive tumors, and the remaining thirteen patients
(35.1 %) presented with TNBC (Table S1). Among these patients, 12
(32.4 %) achieved pCR. The presence of pCR was significantly correlated
with tumor stage and HER2 status, while age, AJCC TNM stage, nodal
status, ER, PR, or histologic grade did not exhibit significant correlations
(Table 1). The logistic analysis unveiled that having a lower tumor stage
and being HER2-positive were favorable indicators for achieving pCR
(Table S2). The differentially expressed genes (DEGs) between pCR and
non-pCR were examined using the Oncomine Immune Response
Research Assay. The data showed that eight genes, KLRK1, IGJ, CD69,
CD40LG, MS4A1, CD1C, KLRB1, and CA4, exhibited higher expression
levels in patients with pCR compared to non-pCR (Figs. 1A and S1). A
heatmap was generated to visualize the expression patterns of these
eight DEGs (Fig. 1B). These DEGs were involved in natural killer cell
activation, B cell marker, T cell receptor signaling, neutrophil, and an­
tigen presentation. To evaluate the predictive performance of these
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Translational Oncology 38 (2023) 101782
DEGs, the area under the receiver operator characteristic curve (AUC)
was calculated. The combined expression of the eight DEGs (referred to
as the 8-gene signature) improved the prediction accuracy of pCR (AUC
= 0.803, Fig. 1C). This 8-gene signature was subsequently validated in
an independent cohort (GSE20194), where it exhibited a higher AUC
compared to individual DEGs (Fig. 1D). These data suggest that the 8-
gene signature was associated with pCR after NACT in breast cancer.
Patients with the 8-gene signature show a favorable outcome
To explore whether pCR-associated DEGs were linked to improved
survival, the RFS of breast cancer patients who received chemotherapy
was analyzed. Data from the KM plotter showed that high-level
expression of seven out of the eight DEGs was associated with longer
RFS. Notably, patients with high levels of the 8-gene signature exhibited
improved RFS (Fig. 2). Likewise, the results were validated using the
METABRIC databases (Fig. S2), suggesting that these DEGs had good
L.-M. Tseng et al. Translational Oncology 38 (2023) 101782

Fig. 3. The expression of eight DEGs is associated with drug response. (A) Data from the ROC plotter database showed the eight DEGs expression in responders and
non-responder breast cancer patients receiving NACT. (B) The dot plot represents the association of IC50 value and gene expression data of cancer cell lines in the
Cancer Therapeutics Response Portal.

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L.-M. Tseng et al.
Fig. 4. Downregulation of eight DEGs in breast cancer tissues. The data of eight DEGs transcript (RNA-Seq by Expectation-Maximization; RSEM) from patients with
breast cancer was selected and analyzed from the TCGA database.
prognostic value. These data indicate that a high expression level of the
8-gene signature was associated with an improved outcome for breast
cancer patients receiving chemotherapy.
The 8-gene signature is linked to drug response
To further study the relationship between DEGs and drug response,
data from the public database ROC plotter was examined. The results
revealed that patients who responded to NACT had higher levels of IGJ,
CD69, and MS4A1 (Fig. 3A). Moreover, drug sensitivity (IC50 values) and
gene expression profiling data of cancer cell lines were depicted by
Spearman correlation. The expression of six out of the eight DEGs was
negatively correlated with drug response. High expression of these DEGs
was sensitive to the drugs including those commonly employed in NACT
regimens for breast cancer, such as taxanes and doxorubicin (Fig. 3B).
The DEGs are downregulated in breast tumor tissues and are correlated
with immune cell infiltrations
To analyze the expression profiles of the eight DEGs in breast cancer,
we examined their transcript levels in the TCGA databases. The
expression of these DEGs, with the exception of MS4A1, was signifi­
cantly downregulated in breast tumor tissues compared to normal tis­
sues (Fig. 4). Next, we used the TIMER database to investigate the
relationship between the DEGs and immune cells. The expression levels
of all eight DEGs were inversely correlated with tumor purity. Among
these DEGs, KLRK1, IGJ, CD69, CD40LG, MS4A1, CD1C, and KLRB1
showed positive correlations with the degree of B cell, CD8+ T cell,
CD4+ T cell, macrophage, neutrophil, and dendritic cell (DC) in­
filtrations (Fig. 5). In addition, pathway analysis revealed that the eight
DEGs were mainly involved in inhibiting the ER and androgen receptor
pathways, while also activating processes such as apoptosis and
epithelial-mesenchymal transition (Fig. S3). Taken together, these re­
sults suggest that the 8-gene signature is not only associated with patient
response to NACT but could also serve as a potential prognostic marker
for breast cancer.
Discussion
Multiple omics technologies provide a powerful tool for identifying
molecular biomarkers in both tumor tissues and liquid biopsies.
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Translational Oncology 38 (2023) 101782
Genomic, transcriptomic, and proteomic markers hold the potential to
enable personalized medicine and enhance the effectiveness of breast
cancer treatment [28]. The development of next-generation sequen­
cing-based platforms has provided a powerful analytical tool to explore
the intricacies of the tumor microenvironment and immunology. This
enables the identification of changes and low-expressing genes in breast
tumor samples. The available genomic tests could serve in determining
the benefit of NACT in ER+/HER2- breast cancer patients. However, the
scope of test panels in the Asian region might be limited due to dis­
crepancies in incidence, mortality rates, epidemiology, and molecular
profiles of breast cancer between Asian and Western populations [29].
Our findings indicate an association between an 8-gene signature and
NACT-induced pCR in Asian populations. This 8-gene signature dem­
onstrates the potential for establishing a novel gene panel as a prog­
nostic biomarker for NACT. Subsequently this biomarker might be
developed as a companion or complimentary assay for NACT for breast
cancer. Furthermore, there exists a potential for future research to
investigate the differences in this 8-gene panel across various ethnicities.
In this study, our data showed an overall pCR rate of 32.4 %.
Notably, patients displaying HER2 expression and those with a lower
tumor stage exhibited an association with pCR. To delve into the intri­
cate dynamics of tumor-immune interactions and NACT, we employed
the Immune Response Research Assay. The outcome demonstrated a
noteworthy linkage between an 8-gene signature and pCR across both
the training cohort (VGH-TAYLOR) and the validation cohort
(GSE20194). Patients exhibiting elevated levels of the 8-gene signature
exhibited improved RFS. Several studies have also indicated that
achieving pCR after NACT is associated with improved survival [4,10,
11,30]. We applied several approaches to explore the potential role of
eight DEGs in breast cancer. Our data showed that these DEGs are
associated with drug response and exhibit downregulation within breast
tumor tissues. Furthermore, a positive correlation between the DEGs and
immune cell infiltrations in breast cancer was observed. However, the
modest sample size and the lack of survival outcome data within the
VGH-TAYLOR cohort are limitations of the current study.
Both KLRK1 and KLRB1 are expressed on natural killer cells and
belong to the C-type lectin superfamily. KLRK1, also known as NKG2D,
functions as an activating receptor that gets induced by cellular stresses,
such as viral infections and tumors. Several studies have demonstrated
that KLRK1 mediates anti-tumor functions in various types of cancer
[31,32]. However, a paradoxical prognostic role has been reported for
L.-M. Tseng et al. Translational Oncology 38 (2023) 101782

Fig. 5. The correlation between eight DEGs and the infiltration levels of immune cells. The correlations of the expression of eight DEGs with tumor purity and the
infiltration levels of B cells, CD8+ T cells, CD4+ T cells, macrophages, neutrophils, and DCs were represented.

KLRK1 ligands [33,34]. Previous studies indicate that KLRB1 could Elevated levels of IGJ (joining chain of multimeric IgA and IgM) tran­
predict better survival in several types of cancer, including breast can­ scripts could serve as a prognostic biomarker in breast cancer [36,37].
cer. KLRB1 expression is correlated with T cell infiltration and immune CD40LG, primarily expressed on activated T cells, regulates the function
checkpoints, suggesting its potential role in immunotherapy [35]. of B lymphocytes by engaging CD40. CD40LG demonstrates a

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L.-M. Tseng et al.
tumor-suppressing role and is associated with a favorable prognosis in
breast cancer [38,39]. MS4A1 encodes the B cell-specific membrane
protein CD20 and plays a crucial role in B cell development and dif­
ferentiation. Recent studies indicate that the presence of CD20 TILs is
associated with improved survival and a higher rate of pCR in inflam­
matory breast cancer [40,41]. CD1C is a member of the CD1 family of
major histocompatibility-like glycoproteins, expressed on the surface of
antigen presenting cells. CD1C-positive DCs mediate the response to
chemotherapeutic-induced immunogenic cell death [42]. Clinical trials
are currently underway to investigate the combination of CD1C-positive
DCs with immunotherapy [43]. CA4, a member of the carbonic anhy­
drase family, catalyzes the reversible hydration of CO2 and dehydration
of HCO−3 , thereby participating in acid/base regulation. Previous studies
have demonstrated that CA4 acts as a tumor suppressor in colon and
gastric cancer [44,45]. However, the role of CA4 in breast cancer re­
mains unclear, despite alteration in acid/base homeostasis being linked
to tumor progression. Unlike the aforementioned seven genes,
CD69-deficiency and anti-CD69 antibody have been shown to exhibit
anti-tumor activity. Studies in CD69-deficient breast cancer models have
revealed reduced tumor growth and metastasis, along with elevated TILs
[46]. CD69 serves as a marker of early leukocyte activation and plays an
essential role in immune response. CD69 is expressed on tissue resident
memory T cells and controls effector T cell retention [47]. Further
investigation is needed to explore the crosstalk between these DEGs in
anti-tumor immunity.
The association between alterations in immune response and prog­
nosis after NACT for breast cancer remains ambiguous. It has been
shown that a combined immune signature is more effective in predicting
pCR than a single biomarker [48]. A higher ratio of T cells to macro­
phages and closer spatial proximity of T cells to tumor cells are associ­
ated with pCR in TNBC [49]. In particular, a randomized phase 3 trial
(IMpassion031) reported that the anti-PD-L1 antibody atezolizumab in
combination with nab-paclitaxel and anthracycline-based chemo­
therapy improves pCR in the NACT setting [50]. Increasing evidence
suggests that the immune profile can predict the response to NACT. In
recent years, immune-related gene signatures have gained considerable
attention in predicting NACT in breast cancer. These signatures
comprise sets of genes associated with immune system activation or
modulation, which was developed to predict pCR to NACT. This classi­
fier showed strong predictive ability for pCR, aiding treatment decision
making and tailored personalized therapy. In TNBC, the status of pCR
versus non-pCR not only forecasts future prognostication but also in­
dicates the application of escalating adjuvant therapy if residual tumors
were to be found in breast or axilla. Overall, immune-related gene sig­
natures provide valuable insights into NACT response prediction, aiding
in treatment decision-making, prognosis estimation, and the develop­
ment of tailored therapeutic strategies.
In summary, our analysis suggests that breast cancer patients with
pCR have higher baseline gene expressions of KLRK1, IGJ, CD69,
CD40LG, MS4A1, CD1C, KLRB1, and CA4 before NACT. The 8-gene
signature is linked to better outcome for patients receiving NACT
regardless of breast cancer subtypes.
Ethics approval and consent to participate
The whole study protocol was reviewed and approved by Institu­
tional Review Board of Taipei Veterans General Hospital (2018-09-
007A). This trial was retrospectively registered (NCT04626440) on Nov
2, 2020.
Consent for publication
Not applicable.
8
Translational Oncology 38 (2023) 101782
Data availability
All data generated or analysed during this study are included in this
published article and its supplementary information files.
Funding
This research is funded from Yong-Lin Healthcare Foundation
(SINO–CANCER project) under the clinical study protocol No.
QCR18002, the National Science and Technology Council, Taiwan
(NSTC 112-2314-B-075-027-MY3), the Ministry of Science and Tech­
nology, Taiwan (MOST 109-2628-B-075-012; MOST 110-2628-B-075-
006; MOST 111-2628-B-075-018), Taipei Veterans General Hospital
(V111C-009; V112C-021), the Taipei Veterans General Hospital—Na­
tional Taiwan University Hospital Joint Research Program (VN110-11;
VN111-06), Dr. Morris Chang (ABMRD002), Melissa Lee Cancer Foun­
dation (MLCF_V112_B11201), and Teh-Tzer Study Group for Human
Medical Research Foundation. The funding sources were not involved in
study design nor manuscript writing.
CRediT authorship contribution statement
Ling-Ming Tseng: Conceptualization, Supervision, Project admin­
istration, Funding acquisition, Investigation, Writing – review & editing.
Chi-Cheng Huang: Formal analysis, Investigation, Resources, Data
curation, Writing – review & editing. Yi-Fang Tsai: Formal analysis,
Investigation, Resources. Ji-Lin Chen: Formal analysis, Data curation,
Writing – original draft. Ta-Chung Chao: Formal analysis, Investiga­
tion, Resources. Jiun-I Lai: Formal analysis, Investigation, Resources.
Pei-Ju Lien: Resources, Data curation, Project administration. Yen-Shu
Lin: Formal analysis, Investigation, Resources. Chin-Jung Feng: Formal
analysis, Investigation, Resources. Yen-Jen Chen: Formal analysis,
Investigation, Resources. Jen-Hwey Chiu: Formal analysis, Resources.
Chih-Yi Hsu: Formal analysis, Investigation, Resources. Chun-Yu Liu:
Conceptualization, Supervision, Project administration, Funding acqui­
sition, Investigation, Writing – review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgments
The authors are grateful to the patients at Taipei Veterans General
Hospital, who provided contributions to enable this research project.
The laboratory works were completed using facilities from Medical
Science & Technology Building of Taipei Veterans General Hospital.
Supplementary materials
Supplementary material associated with this article can be found, in
the online version, at doi:10.1016/j.tranon.2023.101782.
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