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14 18F-Fluciclovine
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Radiolabeled FAPI
https://t.me/med1917
AndreaCimini, MariaRicci, LauraTravascio,
HabibollahDadgar, HosseinArabi, FabioCusella,
NasimNorouzbeigi, HabibZaidi,
andBatoolAlbalooshi
15
15.1 Synthesis
Radiolabeled broblast activation protein inhibitors (FAPIs) are quinoline-based radiopharmaceuticals [1]. In literature, a series of FAPIs have
been reported: the rst variants of FAPI (FAPI-01
and FAPI-02) were documented in 2018: from a
broblast activation protein (FAP)-specic inhibitor, FAPI-01 was created with an organotin stannylated precursor, prepared with a
palladium-catalyzed bromine/tin exchange;
The authors declare they have obtained permission for any
previously published material used in their chapter.
A. Cimini (*) · F. Cusella
Nuclear Medicine Unit, St. Salvatore Hospital,
L’Aquila, Italy
M. Ricci
Nuclear Medicine Unit, Cardarelli Hospital,
Campobasso, Italy
L. Travascio
UOC Nuclear Medicine, P.O.Pescara Santo Spirito,
Pescara, Italy
H. Dadgar · N. Norouzbeigi
Razavi Cancer Research Center, Imam Reza
International University, Mashhad, Iran
H. Arabi
Division of Nuclear Medicine and Molecular
Imaging, Geneva University Hospital,
Geneva, Switzerland
Hossein.Arabi@unige.ch
e-mail:
FAPI-02 was synthesized starting from the same
FAP-specic inhibitor [1].
Other noteworthy variants are represented by
FAPI-04 and FAPI-46. The synthesis of FAPI-04
was reported in 2018, starting with the demethylation of 6-hydroxyquinolie-4-carboxylic acid
with a subsequent etherication, followed by an
amination with 1-tert-butoxycarbonylpiperazine;
nal steps of the synthesis foresee a coupling
between carboxylic acid and glycyl-prolyl and an
amine acylation by DOTA-PNP [2].
FAPI-46 synthesis was documented in 2019,
in which the tert-butyl 6-bromoquinoline-4carboxylate is coupled to the linker reagent by
means of a palladium-catalyzed reaction [2].
H. Zaidi
Division of Nuclear Medicine and Molecular
Imaging, Geneva University Hospital,
Geneva, Switzerland
Geneva University, Geneva, Switzerland
Department of Nuclear Medicine and Molecular
Imaging, University of Groningen, University
Medical Center Groningen, Groningen, Netherlands
Department of Nuclear Medicine, University of
Southern Denmark, Odense, Denmark
Habib.Zaidi@unige.ch
e-mail:
B. Albalooshi
Dubai Nuclear Medicine & Molecular Imaging
Center, Dubai Academic Health Corporation,
Dubai, UAE
bealbalooshi@dha.gov.ae
e-mail:
© The Author(s), under exclusive license to Springer Nature Switzerland AG 2024
F. Calabria, O. Schillaci (eds.), Radiopharmaceuticals,
https://doi.org/10.1007/978-3-031-54196-4_15
259

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A. Cimini et al.
A palladium-catalyzed coupling reaction is
also performed in the synthesis of FAPI-39,
FAPI-40, FAPI-41, FAPI-53, and FAPI-55,
involving the individual linker reagent and tertbutyl 6-bromoquinoline-4-carboxylate [2].
It is important to underline that the use of
DOTA as chelator in radiolabeled FAPIs allows
to incorporate isotopes for diagnostic (68Ga) or
therapeutic purposes (such as 177Lu, 225Ac, or
90Y), offering the possibility of theragnostic
approaches.
An attractive alternative for diagnostic purposes is represented by 18F-labeled FAPIradiotracers, such as 18F-FAPI-74: aluminum
uoride-NOTA complexes and
6- ouronicotineamides are involved in the synthesis and the uorination of this radiopharmaceutical [3].
15.2 Pharmacokinetics
FAP is a type II transmembrane glycoprotein
(belonging to the dipeptidyl peptidase 4 family)
overexpressed on the surface of cancer- associated
broblasts (CAFs) and tumor-associated macrophages (FAP is overexpressed in more than 90%
of epithelial cancers and it is insignicantly
expressed in normal tissues), and consequently, it
represents an attractive target for diagnostic and
therapeutic purposes in nuclear medicine [1, 4,
5]. Moreover, a high expression of FAP has been
demonstrated in inammatory conditions and
brosis [
with subsequent uptake and a rapid internalization in the cell [4, 5]; furthermore, these radiopharmaceuticals have fast clearance from the
body (they are predominantly excreted by the
kidneys) and a rapid accumulation at tumor sites
(10min after the injection) allowing a fast imaging with high contrast in tumors [2, 5]. It is
important to underline that cellular uptake and
retention are different between FAPI variants: for
example, in tumor cells, FAPI-02 has a greater
washout in comparison to FAPI-04 (FAPI-02
retention decreases by 75% in tumor cells from 1
to 3 h after administration, while FAPI-04
1].
Radiolabeled FAPIs specically bind FAP,
decreases by 50%) [2]; variants such as FAPI-46,
FAPI-55, FAPI-36, and FAPI-21 have higher
uptake in tumor cells than FAPI-04, due to a
modication of the linker region that improves
their pharmacokinetic properties. In this context,
FAPI-46 has high tumor-to-organ ratios, resulting in an optimal image contrast for PET imaging
[2].
18F-labeled FAPI-radiotracers such as
18F-FAPI-74 have high uptake in tumor cells, as
revealed in study by Toms et al., making them
attractive for PET imaging [6].
15.3 Physiological Distribution
FAP is a type II serine protease belonging to the
dipeptidyl peptidase 4 family, showing both
dipeptidyl peptidase and endopeptidase activity,
involved in the tumor angiogenesis via cleavage,
among others, of collagen I [7] and in matrix
remodeling of the tumor microenvironment
enabling invasion and migration of tumor cell
[8]. As well as being a cell surface protein, FAP
also exists in a soluble form in human plasma,
truncated of the transmembrane domain, which
converts alpha2-antiplasmin into a more potent
inhibitor of plasmin (LEE k, antiplasmin- cleaving
enzyme blood 2006), promoting scar formation
in adult tissues during wound healing, such as
chronic inammation, after heart infarction and
in liver and lung brosis [7]. Healthy adult tissues, however, show no or very low levels of FAP
expression, except placenta and specic areas of
pancreas, smooth muscles, liver and gallbladder,
kidney and urinary bladder, and cervix and uterus
[
9].
Several variants of molecules targeting FAP,
i.e., FAP-inhibitors such as FAPi-02, FAPi-04,
FAPi-46, FAPi-74, and FAPi-2286, have been
synthetized so far and characterized in mixed
population of different cancers reecting FAP
physiological distribution [10–13].
Biodistribution was assessed by circular regions
of interest (ROIs) in tissues not affected by primary tumor or metastasis and quantied by mean
and maximum standardized uptake values at several time-points (SUVmean and SUVmax)

15 Radiolabeled FAPI
https://t.me/med1917
261
[13–15], eventually comparing FAPi and FDG
SUV values [10].
SUVmax of FAPi was low in brain and lung
parenchyma, myocardial wall, parotid glands,
normal liver, gastrointestinal tract, spleen, and
bone marrow. Breast and pancreas healthy tissues
showed low to mild FAPi uptake, with breast
accumulation depending on the hormonal status.
Conversely, thyroid, oral mucosa, esophagus,
muscles, and blood pool exhibit only mild physiological uptake. Healthy kidney cortex shows
mild to moderate uptake, submandibular salivary
glands moderate to high physiologic accumulation of FAPi tracers. Intense activity is described
in the urinary tract, due to kidneys as the main
excretory organs, and some uptake in the gallbladder and common bile duct, implying elimination via hepatobiliary system as well [16].
Therefore, tumor-to-background ratios in most
normal organs were equal to that of F18FDG.However, high uptake of FAPi was reported
in the uterus, higher in pre-menopausal than in
post-menopausal status (average SUVmax: 11.7
(n=12) vs 3.0 (n =68)) [17]. Finally, reactive
lymph nodes may show FAPi uptake, with SUV
overlapping metastatic nodes SUV values [18].
However, as previously reported, FAP is also
seen in benign tumors, brotic, granulomatosis,
scarring/wound, degenerative and inammatory
disease that can make interpreting FAPI-PET
ndings tricky [18, 19] unless comorbidities and
pitfalls are well-known.
15.4 Clinical Indications
The tumor stroma, which accounts for a large
part of the tumor mass, represents an attractive
target for the delivery of diagnostic (and therapeutic) compounds and, therefore, FAPIs radiopharmaceuticals are emerging as promising tools
in molecular imaging eld, representing appealing targets for therapeutic management and
molecular imaging applications.
FAP is expressed by CAFs, key players in the
multicellular, stromal-dependent alterations promoting cancerous growth [20]. CAFs are present
in more than 90% of epithelial carcinomas [1]
and are among the most abundant components of
the tumor microenvironment in solid tumors
21].
[
Therefore, FAPIs demonstrated highly promising results in previously conducted studies
regarding various tumor entities [17] and in several clinical and research settings [22].
FAPI-PET imaging has opened a new chapter
in diagnostic molecular imaging in oncological
and non-oncological diseases and, even if the
clinical role and indications of FAPI-PET are not
fully established yet, FAPI imaging could potentially replace the FDG-PET scans for different
conditions (oncological and non-oncological)
[11, 23]. These inhibitors have low background
uptake in the brain, liver, and oral/pharyngeal
mucosa and show good contrast between the
tumor and background, which makes up for the
lack of uorodeoxyglucose in the diagnosis of
digestive system tumors [24]. However, further
multicenter trials are needed for a better denition of FAPI imaging role in clinical and research
settings. However, FAP expression is difcult to
detect in non-diseased adult organs, but is greatly
upregulated in sites of tissue remodeling, which
include oncological and non-oncological conditions such as liver brosis, lung brosis, atherosclerosis, arthritis, and embryonic tissues [7].
15.4.1 Oncological Indications
FAPI-PET imaging has been tested in several
kinds of tumor, in consideration of the overexpression of FAP in multiple oncological tissues.
Most of the existing literature data include a
comparison between the FAPI-PET imaging and
the FDG-PET imaging (dominant in oncology),
to dene the potential role of FAPI imaging in
addition (or in replacement) of the validated
imaging tools. Novel FAPI-PET research data,
especially considering a personalized-approach
oncological management, may open new applications for non-invasive tumor characterization and
staging/restaging, or even radioligand therapy.
Further papers are needed to include the FAPIPET imaging in the clinical practice but, according to a recent meta-analysis focused on this

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A. Cimini et al.
topic [22], FAPI-PET imaging appears particularly promising and a possible replacement/addition of FDG-PET imaging in the following
conditions:
1. Cancers that are well or moderately differentiated and, thus, present a relatively slow
growth and a limited Warburg effect.
2. Tumors located close to structures/organs
with variable physiological/inammatory/
drug induced FDG uptake, such as liver and
gut neoplasms.
3. Tumors in areas with permanently elevated
FDG uptake, such as brain and urinary tract
malignancies.
A recent interesting study quantied and compared the tumor uptake on Ga-FAPI PET/CT of
28 different primary and metastatic tumors, to
identify the most promising indications for future
application [11]. According to the authors, SUV
varied across and within all tumor entities with a
low background uptake in muscle and blood pool
and reported the following:
– The highest average SUV was in sarcoma,
esophageal, breast, cholangiocarcinoma, and
lung cancer.
– The intermediate average SUV was in hepato-
cellular, colorectal, head–neck, ovarian, pan-
creatic, and prostate cancer.
– The lowest FAPI uptake was observed in
pheochromocytoma, renal cell, differentiated
thyroid, adenoid cystic, and gastric cancer
[
11].
However, the FAPI-PET imaging was tested
in several categories of tumors and further papers
are needed to dene appropriate clinical
indications.
The FAPI-PET imaging has been tested in
nasopharyngeal carcinoma (NPC), oral squamous cell carcinoma, and Waldeyer’s tonsillar
ring cancer with promising results [11, 25, 26].
FAPI-PET imaging showed in NPC cases a
higher radiotracer uptake than FDG for primary
tumors, regional lymph nodes, and distant metastases, dening a potential suppletory role to MRI
for T staging and therapy planning [
27, 28], par-
ticularly in the evaluation of skull-base and intracranial invasion, suggesting FAPI hybrid PET/
MR has the potential to serve as a single-step
staging modality for patients with NPC [29].
However, the performance of FAPI-PET seems
less promising for N staging because it detected
fewer positive lymph nodes than FDG-PET [28].
Conversely, a further paper indicated that
Ga-FAPI PET/CT might be more specic than
F-FDG for differentiating reactive lymph nodes
from tumor metastatic lymph nodes, supporting
the hypothesis that the FAPI-PET imaging may
distinguish reacting lymph nodes (FDG-positive/
FAPI-negative lymph nodes) from metastatic
lymph nodes. However, this nding requires validation in future research [30].
In oral squamous cell carcinoma (OSCC),
both FAPI and FDG imaging had comparable
sensitivity and specicity for detecting primary
tumors and cervical lymph node metastases [25,
31], but FAPI-PET appears to reduce the false
positivity seen with FDG-PET for the detection
of neck lymph node metastases [32].
In Waldeyer’s tonsillar ring cancers, FAPIPET with respect to FDG-PET showed a higher
detection rate in primary tumors detection, maybe
for the higher tumor-to-background ratio than
FDG, but a reduced detection rate in metastatic
lymph nodes detection [33].
The FAPI-PET imaging has been tested in differentiated thyroid cancer with thyroglobulin
elevation and negative iodine scintigraphy, demonstrating its usefulness for localizing recurrent
or metastatic lesions [
34], especially in lymph
nodes and pulmonary metastases [35]. In addition, FAPI-PET imaging appears promising in
radioiodine-refractory differentiated thyroid cancer (RR-DTC) patients, especially in metastasis
detection [36]. Further papers tested the FAPIPET imaging also in medullary thyroid carcinoma with promising results [11], especially in
liver metastasis detection [37].
FAPI-PET imaging has been tested in cancers
of unknown primary (CUP) [11, 38] with promis-
ing results. FAPI-PET imaging has also been
tested in patients with inconclusive FDG-PET
ndings, showing a potential complementary role

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in discriminating mass lesions and locating the
primary site of unknown malignancy, but also
modifying tumor staging and detecting suspected
disease recurrence in CUP [38, 39] with a performance in assessing metastases like that of
FDG- PET [38]. However, the detection rate of
lymph node metastases was inferior in further
study in a smaller sample, presumably due to low
FAP expression in small metastases [33].
FAPI-PET imaging demonstrated improved
sensitivity in liver, gastric, and pancreatic cancers
when compared with FDG, while the sensitivities
of both tracers were comparable in colorectal and
esophageal cancers [25].
In gastric cancer, FAPI-derived SUVmax in
primary and metastatic lesions was higher than
the FDG-derived SUVmax with higher tracer
uptake in most primary and metastatic lesions in
PET/CT imaging [40], with a superior detection
rate in primary lesion [41, 42]. A further interesting report showed that 68Ga-FAPI PET/MR outperformed 18F-FDG PET/CT in visualizing the
primary and most metastatic lesions of gastric
cancer and might be a promising method, with
the potential of replacing FDG PET/CT [43], particularly in patients with peritoneal carcinomatosis from gastric cancer [44].
In esophageal cancer, the FAPI-PET/CT
methodology showed favorable tumor-to-background contrast in esophageal cancer and might
provide additional information for target volume
delineation (and help avoid tumor geographic
misses) [
45] and in the detection of metastasis
[46] with a possible role also in the chemotherapy response monitoring [47].
Pancreatic tumors are characterized by intense
stromal desmoplastic reactions surrounding cancer cells, and CAFs are the main actors involved
in this desmoplastic reaction. As expected, FAPIPET shows higher sensitivity in detecting primary tumors, lymph nodes, and metastases than
F-FDG PET/CT in pancreatic cancer [25] and is
superior in terms of TNM staging [48]. However,
the FAPI uptake in tumor-induced pancreatitis
represents a possible pitfall, especially in tumors
located in the head of the pancreas. Nevertheless,
an interesting paper described a dual-time point
FAPI-PET/CT acquisition protocol (1 h early-
point and 3h late-point scans) that may help differentiate pancreatitis from malignancy in
primary and recurrent pancreatic carcinomas
[49].
For liver cancer, including hepatocellular carcinoma and intrahepatic cholangiocarcinoma,
FAPI-PET has been demonstrated to have a
higher sensitivity than FDG in detecting primary
liver tumors (partly attributed to higher tumor
uptake and lower hepatic background uptake as
compared with F-FDG) as well as extrahepatic
metastases [25]. In addition, FAPI uptake is correlated mainly with tumor size in the hepatocellular carcinoma [50]. FAPI-PET correctly
identies primary liver tumors and metastasis
equivalent to the CE-CT and liver MRI, but better
than FDG PET. Therefore, FAPI-PET imaging
may improve tumor staging, recurrence detection, and implementation of necessary treatment
modications [51]. Moreover, it was concluded
that FAPI imaging was superior to FDG imaging
in detection of liver metastases of gastrointestinal
system cancer, especially in cases of negative
FDG-PET imaging [52].
Some reports describe the biodistribution of
FAPI-PET in ovarian and endometrial cancer [11,
53]. Due to high tracer uptake resulting in sharp
contrasts in primary and metastatic lesions and
higher TBR than F-FDG-PET/CT, 68Ga-FAPI
PET/CT presents a promising imaging method
for staging and follow-up of gynecological
tumors. The presence or absence of the menstrual
cycle seems to correlate with FAPI accumulation
in the normal endometrium and breast [
17].
FAPI-PET imaging represents a new step in
breast cancer imaging: FAPI-PET is superior to
FDG PET in detecting the primary tumor in
patients with breast cancer with its high sensitivity, high SUVmax, and high TBR.FAPi imaging
appears also superior to FDG PET in detecting
lymph node, hepatic, bone, and cerebral metastases because it has lower background activity and
higher uptake in subcentimetric lesions [54].
Nevertheless, a study reported that FAPI is more
sensitive than FDG in detecting the early stage of
tumor metastasis but becomes less sensitive at
the late stage of tumor metastasis [55]. However,
FAPI-PET could be utilized as an additional

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complementary imaging to 18F-FDG PET/CT,
especially considering its signicant theragnostic
potential, could become a key element in predicting the pathological response of breast cancer
patients in further research [56]. Moreover, novel
research trends may include the promising FAPIPET/MRI [57] that could maximize both morphological and functional aspects of diagnostic
imaging in this eld. However, those interesting
papers need the conrmation of multicenter trials
to dene the denitive role of this new tool.
As regards lung cancer, both FAPI and FDG
had comparable detection rates for primary
tumors in a cohort of 34 patients [25]. Moreover,
FAPI-42 showed higher SUVmax compared to
FDG in the lymph nodes, pleura, bones, and other
tissue lesions [58, 59]. Interestingly, several case
reports and papers agree on the improved detection rate of FAPI imaging, with respect to FDG,
in brain lesions from lung cancers [60, 61], probably due to the higher TBR compared to FDG,
supporting the potential advantages over FDG for
the primary staging of lung. However, these
reports regard small samples and the results are
heterogeneous, in some cases without signicant
differences over the FDG imaging [62].
Notably, FAPI-PET imaging revealed differences in FAP expression in metastases of lung
cancer, with the highest expression specically in
bone metastases, and thus, may be valuable for
distinguishing different pathological types of
lung cancer [63].
Promising results have been described also in
small sample papers and in case-series in the sarcoma’s diagnostic management. Clearly, several
different tumor entities in this eld have been
evaluated by FAPI-PET imaging, with different
results.
An observation trial has been performed in 47
patients with bone or soft-tissue sarcomas, conrming an association between tumoral FAPIPET uptake intensity and histopathologic FAP
expression in sarcoma patients [
64].
Promising results have been reported also:
– In 45 patients with recurrent soft-tissue sar-
coma, especially in terms of restaging [65].
– In a case report of abdominal leiomyosarcoma
[66].
– In a case with intimal sarcoma of the pulmo-
nary artery [67].
– In a case report of gastric Kaposi sarcoma [68]
in which, beyond the promising role in the
detection of gastric inltration, the FDG
seems more useful in the staging.
– In a case of recurrent undifferentiated pleo-
morphic sarcoma of colon mesentery [69].
– In a case of dermatobrosarcoma protuberans
[70].
– In a case of cardiac angiosarcoma [71] in
which FAPI was not superior to FDG.
– In a case of chondrosarcoma of nasal cavity
[72].
– A case report in bilateral primary angiosar-
coma of the breast [73].
Head-to-head comparison studies of hematological neoplasms are relatively rare compared to studies evaluating solid tumors [25].
Moreover, the results are not heterogeneous.
Detecting FAP expression in lymphoma lesions
may be an alternate method for characterizing
lymphoma proles according to previous work
on 73 patients with Hodgkin and non-Hodgkin
lymphomas [74]. However, a direct comparison with FDG imaging demonstrated FAPI’s
inferior sensitivity and accuracy in a subgroup
of eight patients with hematological neoplasms, including multiple myeloma and lymphoma [75].
Interestingly, some case reports describe an
added value in case of tumors in areas with permanently elevated FDG uptake as primary central nervous system lymphoma [
76], hepatic
lesion of mucosa-associated lymphoid tissue
lymphoma [77], and primary hepatic extranodal
marginal zone lymphoma of mucosa-associated
lymphoid tissue [78].
As regards neuroendocrine tumors (NET),
currently, the real-life clinical impact of these
tracers is still mostly not known; however, the
favorable biodistribution and the possibility to
use new theragnostic pairs may provide novel
diagnostic as well as therapeutic options [79].
However, an interesting paper described that
an integrated approach (by performing both FAPI
and DOTATOC imaging) might serve as a clinical parameter for the assessment of dedifferentia-

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tion and aggressiveness of liver metastases in
patients with NET [80].
The biodistribution of FAPI tracers has been
described also in pheochromocytoma [11], in
renal cell carcinoma [11] and prostate cancer [11]
without performing a comparative study with further modalities.
15.4.2 Non-oncological Indications
As studies on FAPI-PET grow in number and
size, incidental ndings related to non-oncologic
conditions have been increasingly reported.
FAPI-PET uptake has been reported in various
conditions such as benign tumors, brotic, granulomatosis, scarring/wound, degenerative diseases, and inammatory diseases. The knowledge
of physiological and non-oncologic causes of
FAPI uptake is indispensable for accurate FAPI-
PET/CT interpretation and can help appropriate
management of incidental ndings on FAPI-PET/
CT in patients referred for cancer staging indications [81]. In addition, FAPI imaging seems
promising also in
– Liver brosis and cirrhosis [82].
– Crohn’s disease where FAP is signicantly
upregulated in the myobroblasts within the
muscle layer of the strictures but not in myobroblasts from patients with ulcerative colitis
[83].
– Arthritis [82].
– IgG4-related disease [82].
– Cardiovascular disease [82].
15.5 Clinical Cases
See Figs.15.1, 15.2, 15.3 and 15.4.
Fig. 15.1 Representative MIP images of 18F-FDG,
68
Ga-DOTATATE (rst image from right-top), and
68
Ga-FAPI PET/CT scans with different cancer types. From
Habibollah Dadgar et al. Initial clinical experience using
Ga-FAPI-46 PET/CT for detecting various cancer types.
Hell J Nucl Med. 2024;27(2):84–93 with permission

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Fig. 15.2 Known case adenocarcinoma colon (41-yearold female) underwent right hemicolectomy and received
chemotherapy before FDG and FAPI scans. Findings from
this patient: (1) Hypermetabolic peritoneal mass lesion in
the RLQ more compatible with peritoneal seeding. In
addition, there are peritoneal nodularities at the surface of
the liver with FAPI uptake more compatible with peritoneal seeding. (2) Two hypermetabolic right renal artery
lymph nodes regarding patients’ history, more compatible
with metastatic involvement. (3) Lung inltration in the
basal left lung with FDG and FAPI uptake, more compatible with inammatory reaction. (4) Three hypermetabolic
small mass lesions in the right liver lobe (segment 8) more
compatible with metastatic involvement. In FAPI-PET/CT,
the scan showed no abnormal uptake throughout the liver
(no evidence of liver metastatic lesion). (5) Physiological
high uptake of FAPI was noted in the uterus. From
Habibollah Dadgar et al. Initial clinical experience using
Ga-FAPI-46 PET/CT for detecting various cancer types.
Hell J Nucl Med. 2024;27(2):84–93 with permission
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