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Chapter 10
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Immunotherapy inBreast Cancer
JennaHoppenworth andRobertoA.Leon-Ferre
Abstract Breast cancer affects 12% of women worldwide. Although advances in
systemic, surgical, and radiation therapy approaches will cure a large proportion of
women with breast cancer, a signicant subset will experience disease relapse and
die as a result of their disease. Of the three main breast cancer subtypes, triplenegative breast cancer poses a special challenge to clinicians and researchers due to
its aggressive features and the paucity of targetable alterations. Recent progress in
immunotherapy and novel therapeutics has transformed the treatment of this breast
cancer subtype. In this chapter, we introduce the reader to the biology and classication of breast cancer and review the modern multidisciplinary approaches to management, with special emphasis on the incorporation of immunotherapy in the
early-stage and metastatic settings. We also review the emerging role of immunotherapy biomarkers and other modern treatments, including monoclonal antibodies
and antibody-drug conjugates.
Keywords Breast cancer · Risk factors · Breast cancer subtypes · Triple-negative
breast cancer · HER2-negative breast cancer · Early-stage breast cancer ·
Metastatic breast cancer · Immunotherapy · Chemotherapy · PD-L1 ·
KEYNOTE-522 · KEYNOTE-355 · Antibody-drug conjugates · Monoclonal
antibodies · Tumor-inltrating lymphocytes
Introduction
Breast cancer will affect approximately 313,510 people in the Unites States in 2024.
Of those affected with breast cancer in 2024, 42,780 will die as a result of their
breast cancer diagnosis (Siegel etal., 2024). Despite the prevalence, the death rate
has steadily declined since the 1990s, mostly credited to early detection and more
J. Hoppenworth · R. A. Leon-Ferre (*)
Division of Medical Oncology, Mayo Clinic, Rochester, MN, USA
e-mail: leonferre.roberto@mayo.edu
H. Dong, S. N. Markovic (eds.), The Basics of Cancer Immunotherapy,
https://doi.org/10.1007/978-3-031-59475-5_10
125© The Author(s), under exclusive license to Springer Nature Switzerland AG 2024

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effective treatments (Siegel etal., 2024). The recent introduction of immunotherapy
has transformed the way we are treating patients with breast cancer, particularly the
triple-negative breast cancer (TNBC) subtype. This chapter will look at the current
role of immunotherapy strategies in the treatment of localized and of metastatic
breast cancer, with special emphasis on TNBC, as well as novel approaches being
investigated that may be incorporated into the clinical management of breast cancer
in the future.
J. Hoppenworth and R. A. Leon-Ferre
Risk Factors
One in eight women will be diagnosed with breast cancer in their lifetime (Siegel
etal., 2024). The risk of developing breast cancer may increase under certain circumstances, such as older age, history of benign breast disease, dense breast tissue,
and being a carrier for inherited genetic mutations such as BRCA 1/BRCA 2,
PALB2, and others (Antoniou etal., 2003; Rahman etal., 2007); having a family
history of breast or ovarian cancer (Collaborative Group on Hormonal Factors in
Breast Cancer, 2001); prior receipt of hormone replacement therapy; reproductive
factors such as earlier onset of menses, late menopause (Collaborative Group on
Hormonal Factors in Breast Cancer, 2012), older age at rst full-time pregnancy, or
nulliparity (Rosner etal., 1994); postmenopausal obesity (Morimoto etal., 2002);
excessive alcohol use or smoking (Gram etal., 2015; Cao etal., 2015); and prior
radiation to the chest at a young age (Henderson etal., 2010); among others. Riskreducing measures include maintaining a healthy weight, abstaining from the use of
alcohol/tobacco, and limiting estrogen replacement therapy, among others.
Basics ofBreast Cancer
Breast cancer evolves when atypical cells develop and divide in an uncontrolled
manner. Benign breast diseases such as atypical ductal or lobular hyperplasia may
progress into ductal or lobular carcinoma in situ or eventually become invasive diseases (with the potential to spread to distant organs). To decrease the risk of potential spread, invasive cancers may require treatment with chemotherapy. Ductal
carcinoma in situ and lobular carcinoma in situ are noninvasive cancers (without the
potential to spread) and are often referred to as stage 0 cancer. Given that they typically remain localized, chemotherapy is not recommended.
Breast cancers are classied biologically by the presence or absence of several
markers, most importantly estrogen, progesterone, and HER2 (human epidermal
growth factor receptor 2) receptors. Other features such as their appearance under
the microscope (known as histologic subtype), grade (how mature or immature cells
look in a biopsy specimen), and Ki-67 (a marker of cell proliferation) help breast
cancer specialists determine the most effective and appropriate treatment.

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Breast Cancer Subtypes
Breast cancer can be categorized into three major subtypes, according to the presence of three major markers within tumors, as outlined in Fig.10.1 and below:
• Hormone receptor-positive breast cancer: It is characterized by the expression
of the estrogen and/or progesterone receptors. In this subtype, growth is stimu-
lated by estrogen and the activation of its receptor. Antiestrogen therapy is a
major component of the treatment of patients with this tumor subtype. In certain
situations, chemotherapy may also be necessary.
• HER2-positive breast cancer: These tumors are characterized by excessive
amounts of HER2, a normal protein involved in normal cell division. In this
breast cancer subtype, the presence of an abnormally high number of HER2
proteins contributes to excessive growth of cancer cells. To treat these tumors,
therapies targeting the HER2 signaling pathway have been developed. These
drugs are typically combined with chemotherapy.
• Triple-negative breast cancer: These tumors are characterized by absence of the
estrogen and progesterone receptor and lack HER2 overexpression. This subtype
Fig. 10.1 Breast cancer subtypes. The three main breast cancer subtypes are classied according
to the presence or absence of the hormone receptors (estrogen and progesterone receptors) and the
presence or absence of excessive HER2 signaling. Hormone receptor-positive breast cancers are
the least susceptible to the effects of the immune system, while triple-negative breast cancers (lacking all three features) are the most susceptible to the effects of the immune system

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is the most aggressive type of breast cancer. For many years, chemotherapy has
been the main form of treatment. However, this subtype has benetted the most
from the development of immunotherapy.
Each of these subtypes of breast cancer has a different biology and clinical
behavior, requiring different treatment approaches as introduced briey here.
Importantly, immunotherapy approaches have demonstrated the greatest degree of
clinical activity in the TNBC subtype. Given this, we will focus our subsequent
discussions primarily on TNBC.
J. Hoppenworth and R. A. Leon-Ferre
Early-Stage Breast Cancer: Multidisciplinary Approaches
toTreatment
Early-stage breast cancer (breast cancer that is conned to the breast and regional
lymph nodes and has not spread distantly) can be eradicated and treated with the
goal of cure. To achieve this, specialists from different medical disciplines work
together. These include surgical oncologists, radiation oncologists, medical oncologists, geneticists, physical therapists, and plastic surgeons, among others. It is
important to understand the role each discipline plays in the treatment of breast
cancer and the contribution of each treatment modality in cancer eradication and
prevention of recurrence or metastatic disease. Advances in each treatment modality
and in the integration of multidisciplinary care have led to substantial improvements
in recurrence rates and breast cancer survival (Poortmans et al., 2015; Whelan
etal., 2015).
Treatments specically targeting the tumor within the breast and affected lymph
nodes are called “locoregional therapies” and include surgery and radiation therapy
(Fig. 10.2). Medications aiming to treat not only the breast and affected lymph
nodes but also any breast cancer cells that may have spread to the rest of the body
are called “systemic therapies” and are managed by the medical oncologist. Cancer
cells that are not eradicated from the breast can lead to local recurrence (reappearance of tumor in the same breast). Cancer cells that have accessed the circulation or
lymphatic networks can lead to regional recurrence (reappearance of tumor in the
draining lymph nodes) or distant recurrence (spread to other organs, which is often
incurable). Cancer cells that have spread before colonizing organs are often microscopic and not detectable by traditional imaging. As such, the multidisciplinary
team evaluates the risk for potential microscopic metastatic disease and based on
this makes recommendations for the need of different treatment modalities.
Immunotherapy is primarily considered a form of systemic therapy, with their
main goal being to help activate the immune system’s inherent ability to recognize
and eliminate abnormal cells (in this case cells that have become cancerous).
However, certain local immunotherapies have also been developed and are under
investigation (examples of these include injections of medications or immune stimulants into the tumor or lymph nodes).

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Fig. 10.2 Locoregional and systemic therapies for breast cancer. Surgery and radiotherapy are
considered locoregional therapies, while medications including chemotherapy, immunotherapy,
and targeted therapies are considered systemic therapies
How Are theDifferent Treatment Modalities Integrated?
Over the last decade, the multidisciplinary management paradigm of early-stage
breast cancer, particularly for TNBC and HER2-positive breast cancer, has shifted
away from doing surgery rst and systemic therapy after surgery (called “adjuvant”
systemic therapy) to a preference for administering systemic therapy before surgery
(called “neoadjuvant” systemic therapy). This strategy of giving systemic therapy
rst has allowed clinicians to evaluate the sensitivity of the cancer to the treatment
as it is administered and make adjustments as needed. The administration of neoadjuvant chemotherapy has several direct benets to the patient with breast cancer,
including (1) potential for less extensive breast and axillary surgery, in turn leading
to decreased surgical morbidity and complications, given that tumors and lymph
nodes often shrink or even disappear after neoadjuvant chemotherapy (Piltin etal.,
2020; Boughey et al., 2018), allowing surgeons to remove smaller amounts of

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affected tissue; (2) better ability to estimate future risk of recurrence (Cortazar etal.,
2014; Symmans etal., 2007; Boughey etal., 2017; Symmans etal., 2017; Yau etal.,
2022), given that the degree of response of the breast tumor and lymph nodes to
systemic therapy is strongly associated with the subsequent risk of cancer recurrence; (3) opportunity to individualize postoperative treatment decisions, allowing
clinicians to selectively recommend additional medications to patients with more
resistant tumors, and avoiding those medicines in patients with more sensitive
tumors that were eradicated prior to surgery; and (4) more rapid evaluation of newer
drugs, as clinical trials testing new drugs while the tumor has not been removed yet
allows to quickly evaluate their efcacy (Masuda etal., 2017; Tutt etal., 2021a).
In breast cancer, the response to neoadjuvant systemic therapy has repeatedly
been shown to strongly correlate with long-term outcomes, including risk of recurrence and long-term survival (Cortazar etal., 2014; Symmans etal., 2007; Boughey
etal., 2017; Symmans etal., 2017; Yau et al., 2022). Patients whose tumors are
completely eradicated by the end of neoadjuvant systemic therapy (called a pathologic complete response or pCR) enjoy very favorable long-term survival, with
recurrence and mortality rates of <10% (Symmans etal., 2017). On the other hand,
patients whose tumors or parts of their tumors survive after neoadjuvant systemic
therapy (called residual disease) experience higher rates of disease recurrence and
death—with the risk increasing as the amount of residual disease increases (Yau
etal., 2022). This risk can be mitigated by administering additional medications
after surgery for patients who have residual disease.
J. Hoppenworth and R. A. Leon-Ferre
Breast Cancer andtheImmune System
As elaborated throughout this book, immunotherapy has transformed the treatment
landscape and prognosis of aggressive malignancies that previously had limited systemic therapy options. In breast cancer, patients with TNBC have derived the largest
benets from immunotherapy. This is in part due to TNBC tumors being better able
to trigger immune system responses than HER2-positive or hormone receptorpositive breast cancer (Cortes etal., 2020; Loibl etal., 2019; Mittendorf etal., 2020;
Nanda etal., 2020; Schmid etal., 2020). Importantly, TNBC tumors that generate
more robust immune responses generally have better clinical outcomes than TNBC
tumors with weaker immune responses. For example, a measure of the ability of a
tumor to trigger immune responses is the observation of immune system cells such
as lymphocytes invading within the tumor tissues. These immune system cells,
often called “tumor-inltrating lymphocytes” (TILs), have the ability to recognize
cancer cells and—if appropriately stimulated—eradicate them. Compared with
TNBC that contains low levels of TILs, patients with TNBC that contains high levels of TILs have better survival following adjuvant systemic therapy (Loi etal.,
2019), higher rates of pCR following neoadjuvant systemic therapy (Denkert etal.,
2018), and in select cases (such as in stage I TNBC) better survival even without
systemic therapy (Leon-Ferre etal., 2018; Park et al., 2019; de Jong etal., 2022;

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Leon-Ferre etal., 2024). TIL levels are also associated with prognosis and response
to systemic therapy in HER2-positive breast cancer (Loi etal., 2014) but less so in
hormone receptor-positive breast cancer (Criscitiello etal., 2020). As such, TILs
and other features of immune activation are promising biomarkers that may allow
clinicians to optimize systemic therapy for TNBC and possibly HER2-positive
breast cancer in the future.
In the clinic, expression of a protein called programmed death-ligand 1 (PD-L1)
in the tumor microenvironment is currently used to identify patients with metastatic
TNBC who may benet from immunotherapy. This protein can be found on the
surface of tumor cells and in immune cells. When activated and through interactions
with another protein (called programmed cell death protein 1 or PD-1), it sends a
signal to T cells from the immune system to stop attacking the tumor cells, shielding
them from eradication by the immune system. PD-1 and PD-L1 blocking medications effectively release the breaks from the immune system and restore the ability
of T cells to recognize and attack breast cancer tumor cells.
Several studies have shown that in patients with metastatic TNBC, the addition
of immunotherapy to chemotherapy only benets patients whose tumors express
PD-L1, whereas patients with tumors that are negative for PD-L1 do not appear to
derive benet. However, in patients with localized TNBC treated with multimodality approaches, the use of immunotherapy appears to benet patients regardless of
whether their tumors express PD-L1 or not. For this reason, testing for PD-L1
expression is only recommended in the setting of metastatic TNBC and not in earlystage TNBC. However, challenges with this biomarker remain, as there are multiple
assays, methodologies and cutoff points to determine “positivity” versus “negativity” (Gonzalez-Ericsson etal., 2020).
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Chemoimmunotherapy forEarly-Stage Triple-Negative
Breast Cancer
TNBC is more aggressive than other breast cancer subtypes and has historically
been associated with higher and earlier rates of disease recurrence and death (Lin
etal., 2012; Cortes et al., 2022; Bardia et al., 2021a). TNBC represents approximately 15% of all breast cancer cases and more often affects younger women and
racial and ethnic minority groups including Black, Latina, and Indian women (Lin
etal., 2012; Friebel-Klingner etal., 2021; DeSantis etal., 2019; Du, 2022; Kulkarni
etal., 2020). As elaborated previously, modern management of early-stage TNBC
generally favors administration of systemic therapy prior to surgery for several reasons, including the ability to assess the effect of the treatment on the tumor cells
prior to being surgically removed. In TNBC, it has become very clear that attainment of pCR is strongly associated with lower recurrence rates and decreased mortality (Cortazar etal., 2014; Symmans etal., 2007; Boughey etal., 2017; Symmans
etal., 2017; Yau etal., 2022). As such, one of the major goals of systemic therapy

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J. Hoppenworth and R. A. Leon-Ferre
optimization has been to incorporate drugs that can increase pCR and subsequently
improve long-term outcomes. Patients who do not achieve pCR can then be offered
additional systemic therapy to further decrease risk of recurrence (Masuda
etal., 2017).
Until recently, the mainstay of systemic therapy for TNBC was cytotoxic chemotherapy, often including a combination of an anthracycline (such as doxorubicin or
epirubicin, among others) paired with cyclophosphamide and followed by (or preceded by) a taxane (such as paclitaxel or docetaxel). Several studies tested whether
incorporating additional agents could improve the rates of pCR and long-term outcomes. The incorporation of immunotherapy agents (particularly pembrolizumab)
to neoadjuvant chemotherapy has been one of the major advances of the management of early-stage TNBC, leading to improvements both on pCR rates and eventfree survival (the time between treatment and the occurrence of a new “event” such
as recurrence, a new primary cancer or death). The KEYNOTE-522 clinical trial
evaluated the addition of pembrolizumab to a chemotherapy backbone consisting of
carboplatin + paclitaxel followed by an anthracycline and cyclophosphamide
(Schmid etal., 2020). In this trial, 1174 patients with newly diagnosed stage 2 or 3
TNBC were randomly assigned (in a 2:1 fashion) to receive IV pembrolizumab plus
chemotherapy (paclitaxel and carboplatin followed by doxorubicin and cyclophosphamide) versus standard of care with chemotherapy plus placebo. Eligibility
patients had newly diagnosed node-positive TNBC or tumors measuring ≥2 cm
(regardless of nodal involvement). Patients with both PD-L1-positive and PD-L1negative TNBC were eligible (although 84% in the treatment arm were PD-L1 positive). Following surgery, patients received nine additional cycles of adjuvant
pembrolizumab or placebo every three weeks regardless of treatment response.
The incorporation of pembrolizumab to neoadjuvant chemotherapy increased the
proportion of patients achieving a pCR (63%) compared to chemotherapy alone
(56%), leading to an improvement in pCR rates of 7%. Furthermore, patients who
received pembrolizumab displayed an improved event-free survival (85% with
pembrolizumab compared to 77% with chemotherapy alone, an improvement of
8%). While patients who achieved pCR in either treatment arm had very similarly
favorable outcomes (three-year EFS of 94% vs. 93% with or without immunotherapy), patients who had residual disease following treatment with immunotherapy
had improved three-year event-free survival (67%) compared to those who received
chemotherapy alone (57%).
As previously mentioned, PD-L1 expression does not appear to help distinguish
which patients with early-stage TNBC benet from the addition of pembrolizumab
(unlike in the metastatic setting). However, in KEYNOTE-522, patients with TNBC
expressing higher levels of PD-L1 demonstrated higher rates of pCR (both with and
without immunotherapy).
Several outstanding questions remain in the eld regarding the optimal incorporation of immunotherapy for early-stage TNBC. These are being actively research
and include the following:
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