Craniofacial morphologic alteration induced by bone-targeted mutants of FGFR2 causing Apert and Crouzon syndrome

Lee, Kee-Joon,Nah, Hyun-Duck,Tjoa, Stephen T. J.,Park, Young-Chel,Baik, Hyoung-Seon,Yun, Tae-Min,Song, Jin-Wook 대한치과교정학회 2006 대한치과교정학회지 Vol.36 No.4

Objective: Activating mutations in the fibroblast growth factor receptor-2 (FGFR2) have been shown to cause syndromic craniosynostosis such as Apert and Crouzon syndromes. The purpose of this pilot study was to investigate the resultant phenotypes induced by the two distinctive bone-targeted gene constructs of FGFR2, Pro253Arg and Cys278Phe, corresponding to human Apert and Crouzon syndromes respectively. Methods: Wild type and a transgenic mouse model with normal FGFR2 were used as controls to examine the validity of the microinjection. Micro-CT and morphometric analysis on the skull revealed the following results. Results: Both Apert and Crouzon mutants of FGFR2 induced fusion of calvarial sutures and anteroposteriorly constricted facial dimension, with anterior crossbite present only in Apert mice. Apert mice differed from Crouzon mice and transgenic mice with normal FGFR2 in the anterior cranial base flexure and calvarial flexure angle which implies a possible difference in the pathogenesis of the two mutations. In contrast, the transgenic mice with normal FGFR2 displayed normal craniofacial phenotype. Conclusion: Apert and Crouzon mutations appear to lead to genotype-specific phenotypes, possibly causing the distinctive sites and sequence of synostosis in the calvaria and cranial base. The exact function of the altered FGFR2 at each suture needs further investigation. 유전적으로 결정되는 두개안면 기형의 발생 기전을 밝히기 위해 관련된 유전자의 기능 변화에 의한 효과를 이해하는 것이 필수적이다. 섬유아세포성장인자수용체-2 (FGFR2)의 활성형 돌연변이가 어퍼트 및 크루즌 증후군에서의 봉합의 조기유합의 원인이 된다고 알려져 있으나 인류에서는 다양한 개인차가 존재하므로 임상적으로 정의된 두 증후군에서의 유전형-표현형의 상관관계에 대해서는 의문이 제기되어 왔다. 본 연구의 목적은 어퍼트(Pro253Arg)및 크루즌(Cys278Phe) 돌연변이를 갖는 골특이성 FGFR2를 발현하도록 제작된 형질변환 쥐에서 결과적인 표현형의 차이를 분석하여 유전형에 의한 기형 형성의 인과관계를 추정하기 위한 것이다. 유전자 조작을 하지 않은 정상군과 정상 FGFR2 유전자를 가진 군을 대조군으로 하여 육안 관찰 및 micro-CT를 이용한 형태계측학적 방법으로 주로 전방두개 및 전두개저 부위의 이상을 분석하여 다음과 같은 결론을 얻었다. 첫째, 어퍼트 및 크루즌 돌연변이를 갖는 각각의 형질변환 쥐는 두개 봉합의 유합과 전후방적 두부 길이 감소를 공히 보였으나 어퍼트 형질변환 쥐에서만 전치부 반대교합이 나타났다. 또한 어퍼트 개체는 크루즌 개체 및 대조군에 비해 전방두개 및 전두개저 굴곡에 있어서 정상군과 비교해 차이를 나타냈으며 이는 유합을 보이는 봉합의 부위 및 순서에 있어서의 차이에 기인하는 것으로 사료된다. 둘째, 정상 FGFR2 유전자를 주입한 형질변환 쥐는 정상적인 두개안면 형태를 보였다. 이상의 결과를 토대로 어퍼트 및 크루즌 돌연변이는 각각의 유전형에 특이한 두개안면 기형을 유발할 것으로 보이며 정상 FGFR2의 발현 강도보다는 기능의 이상이 두개골 유합과 관련이 있는 것으로 사료된다. 변형된 FGFR2와 각 봉합에서의 기능과의 상관성은 추가 연구가 필요할 것으로 사료된다.

FGF-mediated FGFR signaling 이 두개봉합부의 초기형태발생 및 유지기전에 미치는 영향

남순현,김영진,서경환,김현정,박미현,유현모 大韓小兒齒科學會 1999 大韓小兒齒科學會誌 Vol.26 No.4

두개봉합부의 조기융합으로 알려진 Craniosynostosis는 두개봉합부 주위 조직들 사이의 조화로운 상호 작용이 파괴되었을 때 야기될 수 있다. 흥미롭게도 FGF receptor들, 특히 FGFR2의 point mutation은 여러 가지 형태의 craniosynostosis 증후군과 연관되어 있어, FGFR가 두개봉합부를 포함한 두개골 성장 발달과정에 중요한 유전자임을 시사하고 있다. Mouse 두개봉합부의 초기형태발생시 FGFR 유전자들의 기능을 알아보기위해, in situ hybridization 방법을 이용하여 FGFR2(BEK) 및 골아세포분화의 초기표지자인 osteopontin이, 태생기(E15-18)에서 출생후(P1-P3)까지, 두개골의 시상봉합부에서의 발현양상을 조사하였다. FGFR2(BEK)은 osteogenic front에 강하게 발현되었으며, osteopontin은 parietal bone의 exo-, endocranial부위에서 발현되었으나, parietal bone의 성장가장자리인 osteogenic front에서는 관찰되지 않았다. 두개봉합부에서의 FGF-mediated FGFR signaling의 역할을 좀더 심도깊게 조사 하기위해 E15.5 mouse의 두개골을 이용하여 in vitro 실험을 시행하였다. 흥미롭게도 osteogenic fronts 및 시상봉합부의 간엽조직 중앙에 FGF2 - soaked beads를 점적하여 36시간 기관배양한 결과, bead주위 조직들의 두께 및 세포수가 증가되었으며, osteogenic fronts 상에 FGF4 beads를 올려놓은 경우, 시상두개봉합부 중앙에 점적된 FGF4 beads나 BSA control beads에 비해, 골성장이 촉진되어 시상두개봉합부의 부분적인 소멸을 관찰할 수 있었다. 이와 더불어 FGFR2 beads는 osteopotin 및 Msxl 유전자의 발현을 유도하였다. 이 결과들을 종합해 볼 때, FGF - mediated FGFR signaling이 발육중인 두개골과 두개봉합부에서 세포의 증식과 분화의 균형을 조절하는데 중요한 담당하고 있음을 시사해주고 있으며, 이 과정중 FGF signaling이 osteopontin 및 Msxl 유전자의 발현을 조절하므로써 막내골 성장 및 두개봉합부의 유지기전에 기여할 것으로 사료된다. Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of the interactions between different tissues within the cranial sutures. Interestingly, point mutaions in the genes encoding for the fibroblast growth factor receptors(FGFRs), especially FGFR2, cause various types of human craniosynostosis syndromes. To elucidate the function of these genes in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of FGFR2(BEX) and osteopontin, an early marker of osteogenic differentiation, in the sagittal suture of calvaria during embryonic (E15-E18) and postnatal stage (P1 - P3). FGFR2(BEK) was intensely expressed in the osteogenic fronts, whose cells undergo differentiation into osteoprogenitor cells that ultimately lay down the bone matrix. Osteopontin was expressed throughout the parietal bones excluding the osteogenic fronts, the periphery of the parietal bones. To further examine the role of FGF-mediated FGFR signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of FGF2 soaked beads onto both the osteogenic fronts and mid-mesenchyme of sagittal suture after 36 hours organ culture resulted in the increase of the tissue thickness and cell number around FGF2 beads, moreover FGF4-soaked beads implanted onto the osteogenic fronts stimulated suture closure due to an accelerated bone growth, compared to FGF4 beads placed onto mid-mesenchyme of sagittal suture and BSA control beads. In addition FGF2 induced the ectopic expression of osteopontin and Msxl genes. Taken together, these data indicate that FGF-mediated FGFR signaling has a important role in regulating the cranial bone growth and maintenance of cranial suture, and suggest that FGF-mediated FGFR signaling is involved in regulating the balance between the cell proliferation and differentiation through inducing the expression of osteopontin and Msxl genes.

Association Study of Fibroblast Growth Factor 2 and Fibroblast Growth Factor Receptors Gene Polymorphism in Korean Ossification of the Posterior Longitudinal Ligament Patients

Jun, Jae-Kyun,Kim, Sung-Min The Korean Neurosurgical Society 2012 Journal of Korean neurosurgical society Vol.52 No.1

Objective : The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of fibroblast growth factor (FGF) 2 gene and fibroblast growth factor receptor (FGFR) genes are associated with ossification of the posterior longitudinal ligament (OPLL). Methods : A total of 157 patients with OPLL and 222 controls were recruited for a case control association study investigating the relationship between SNPs of FGF2, FGFR1, FGFR2 and OPLL. To identify the association among polymorphisms of FGF2 gene, FGFR1, FGFR2 genes and OPLL, the authors genotyped 9 SNPs of the genes (FGF2 : rs1476217, rs308395, rs308397, and rs3747676; FGFR1 : rs13317 and rs2467531; FGFR2 : rs755793, rs1047100, and rs3135831) using direct sequencing method. SNPs data were analyzed using the SNPStats, SNPAnalyzer, Haploview, and Helixtree programs. Results : Of the SNPs, a SNP (rs13317) in FGFR1 was significantly associated with the susceptibility of OPLL in the codominant (odds ratio=1.35, 95% confidence interval=1.01-1.81, p=0.048) and recessive model (odds ratio=2.00, 95% confidence interval=1.11-3.59, p=0.020). The analysis adjusted for associated condition showed that the SNP of rs1476217 (p=0.03), rs3747676 (p=0.01) polymorphisms in the FGF2 were associated with diffuse idiopathic skeletal hyperostosis (DISH) and rs1476217 (p=0.01) in the FGF2 was associated with ossification of the ligament flavum (OLF). Conclusion : The results of the present study revealed that an FGFR1 SNP was significantly associated with OPLL and that a SNP in FGF2 was associated with conditions that were comorbid with OPLL (DISH and OLF).

Generation of a transgenic mouse model to study cranial suture developement : Apert syndrome 어퍼트 신드롬

Lee, Kee-Joon,Chootima Ratisoontorn,Baik, Hyoung-Seon,Park, Young-Chel,Park, Kwang-Kyun,Nah, Hyun-Duck 대한치과교정학회 2003 대한치과교정학회지 Vol.33 No.6

악안면 구조의 형태와 기능은 대개 유전자 정보에 의해 결정된다. 분자생물학의 발달로 인해 정상 성장과 형태 형성에 중요한 유전자에 대한 정보가 밝혀지고 있고 이는 현대 두개안면 생물학의 근간이 되고 있다. 밝혀진 사실들 중 주목할 만한 것은 섬유아세포 성장인자2 (FCFR2)에서의 특이한 돌연변이가 어퍼트 증후군 (Apert syndrome) 의 발생과 관련이 있다는 것이다. 어퍼트 증후군은 두개 관상봉합의 조기 유합과 사지의 기형으로 특징지워진다. 그 중 특히 두개골 유합증의 병인과 형성기전을 연구하기 위해 본 연구에서 유전자 변환기법을 시도하여 어퍼트 증후군의 유발인자로 알려진 FCFR2에서의 단일 아미노산 치환 돌연변이를 재연한 인위 유전자구조물을 제작하고 이를 미세주입법으로 쥐의수정란에 삽입하여 형질변환 쥐를 제작하였다. 본 연구에서는 전체 조직이 아닌 골조직에서 특이하게 활성화되는 전사촉진자(promoter, 제Ⅰ형 교원질 유전자의 전사촉진자) 를 이용하여 골조직에서만 돌연변이 유전자의 발현을 재현함으로써 이 시도가 쥐에서 두개골유합증을 유발하는지 검증하고자 하였다. 초기 표현형 분석을 통해 어퍼트 환자에서 기대되는 두개골 유합증을 확인하였다. 또한 본 연구에서 삽입된 변환유전자가 원활히 돌연변이 단백질을 생산하고 기능의 증가를 보임을 확인하였다. 이러한 동물 모델을 이용함으로써 이제 정상적 혹은 비정상적 두개골 및 봉합 발육에서의 FCFR2의 역할을 연구하는 것이 가능하리라 사료된다. The form and function of the craniofacial structure critically depend on genetic information. With recent advances in the molecular technology, genes that are important for normal growth and morphogenesis of the craniofacial skeleton are being rapidly uncovered, shaping up modern craniofacial biology. One of them is fibroblast growth factor receptor 2 (FGFR2). Specific point mutations in the FGFR2 gene have been linked to Apert syndrome, which is characterized by premature closure of cranial sutures and craniofacial anomalies as well as limb deformities. To study pathogenic mechanisms underlying craniosynostosis phenotype of Apert syndrome, we used a transgenic approach; an FGFR2 minigene construct containing an Apert mutation (a point mutation that substitute proline at the position 253 to arginine; P253R) was introduced into fertilized mouse germ cells by DNA microinjection. The injected cells were then allowed to develop into transgenic mice. We used a bone-specific promoter (a DNA fragment from the type I collagen gene) to confine the expression of mutant FGFR2 gene to the bone tissue, and asked whether expression of mutant FGFR2 in bone is sufficient to cause the craniosynostosis phenotype in mice. Initial characterization of these mice shows prematurely closed cranial sutures with facial deformities expected from Apert patients. We also demonstrate that the transgene produces mutant FGFR2 protein with increased functional activities. Having this useful mouse model, we now can ask questions regarding the role of FGFR2 in normal and abnormal development of cranial bones and sutures.

Functional characterization of a novel FGFR2 mutation, E731K, in craniosynostosis

Park, Jounghyen,Park, Ok‐,Jin,Yoon, Won‐,Joon,Kim, Hyun‐,Jung,Choi, Kang‐,Young,Cho, Tae‐,Joon,Ryoo, Hyun‐,Mo Wiley Subscription Services, Inc., A Wiley Company 2012 Journal of cellular biochemistry Vol.113 No.2

<P><B>Abstract</B></P><P>Craniosynostosis is a condition in which some or all of the sutures in the skull of an infant close prematurely. Fibroblast growth factor receptor 2 (FGFR2) mutations are a well‐known cause of craniosynostosis. Many syndromes that comprise craniosynostosis, such as Apert syndrome, Crouzon syndrome, and Pfeiffer syndrome, have one of the phenotypes that have been reported in FGFR2 mutant patients. FGFRs have been reported in four types (FGFR1–4), and upon binding with FGF ligands, signal transduction occurs inside of cells. Activated FGFR stimulates an osteogenic master transcription factor, Runx2, through the MAP kinase and PKC pathways. We obtained a genetic analysis of six Korean patients who have craniosynostosis as a phenotype. All of the patients had at least one mutation in the FGFR2 gene; five of those mutations have already been reported elsewhere, while one mutation is novel and was hypothesized to lead to Apert syndrome. In this study, we reported and functionally analyzed a novel mutation of the FGFR2 gene found in a craniosynostosis patient, E731K. The mutation is in the 2nd tyrosine kinase domain in the C‐terminal cytoplasmic region of the molecule. The mutation caused an enhanced phosphorylation of the FGFR2<SUP>E731K</SUP> and ERK‐MAP kinase, the stimulation of transcriptional activity of Runx2, and consequently, the enhancement of osteogenic marker gene expression. We conclude that the substitution of E731K in FGFR2 is a novel mutation that resulted in a constitutive activation of the receptor and ultimately resulted in premature suture obliteration. J. Cell. Biochem. 113: 457–464, 2012. © 2011 Wiley Periodicals, Inc.</P>

<i>FGFR2</i> Assessment in Gastric Cancer Using Quantitative Real-Time Polymerase Chain Reaction, Fluorescent In Situ Hybridization, and Immunohistochemistry

Park, Young Soo,Na, Young-Soon,Ryu, Min-Hee,Lee, Chae-Won,Park, Hye Jin,Lee, Ju-Kyung,Park, Sook Ryun,Ryoo, Baek-Yeol,Kang, Yoon-Koo American Society for Clinical Pathology 2015 American journal of clinical pathology Vol.143 No.6

<P><B>Objectives:</B></P><P>Fibroblast growth factor receptor 2 <I>(FGFR2)</I> amplification has been reported to be a target for treatment in gastric cancer. However, an optimal tissue source and method for evaluating <I>FGFR2</I> have yet to be established.</P><P><B>Methods:</B></P><P>Copy numbers were compared by quantitative polymerase chain reaction (qPCR) using frozen vs formalin-fixed, paraffin-embedded (FFPE) tissue and biopsy vs surgical specimens. We correlated the results of qPCR and immunohistochemistry (IHC) with fluorescence in situ hybridization (FISH) using stage IV gastric cancer biopsy specimens and validated the results in surgical specimens.</P><P><B>Results:</B></P><P>FFPE tissues were suitable for qPCR, and biopsy specimens were equivalent to or better than surgical specimens. qPCR and IHC results exhibited an excellent correlation with FISH at eight or more copies by qPCR in any kind of tissue, 5% or more by IHC in biopsy specimens, and 10% or more by IHC in surgical specimens. <I>FGFR2</I> amplification was 6.6% in stage IV gastric cancers, and 42% of these showed heterogeneous amplification and overexpression. IHC indicated a good correlation with FISH even in the heterogeneous cases.</P><P><B>Conclusions:</B></P><P>FFPE biopsy tissues are an adequate source for <I>FGFR2</I> evaluation in gastric carcinomas, and a qPCR-based copy number assay can be used for screening. IHC is also a valid and practical method for evaluating <I>FGFR2</I>, considering frequent heterogeneity.</P>

Association of rs1219648 in FGFR2 and rs1042522 in TP53 with Premenopausal Breast Cancer in an Iranian Azeri Population

Saadatian, Zahra,Gharesouran, Jalal,Ghojazadeh, Morteza,Ghohari-Lasaki, Sahar,Tarkesh-Esfahani, Najime,Ardebili, Seyyed Mojtaba Mohaddes Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.18

Breast cancer is the most common cancer among women in the world. In Iran, the incidence of breast cancer is on the increase. We here studied the association of rs1219648 in FGFR2 and rs1042522 in TP53 and their interaction in development of early onset sporadic breast cancer in Iranian Azeri population to evaluate epistatic effects on the risk of mammary neoplasia. We genotyped the two polymorphisms in 100 women with early onset breast cancer and 100 healthy women by PCR-RFLP. Allele frequency differences were tested using $chi^2$-test with 95% confident intervals. Our results indicated a statistically significant association (p<0.05) between rs1219648, but not rs1042522, and risk of breast cancer. We also found that the combination of FGFR2 major genotype and TP53 hetero genotype had protective effects against breast cancer, while the hetero allele of FGFR2 in combination with the minor genotype of TP53 was associated with a high risk. This study revealed an important crosstalk between two polymorphisms in FGFR2 and TP53 in development of breast cancer. These candidates risk variants should be further evaluated in studies with a larger sample size.

FGFR2 유전자의 8번째 엑손부위의 P253R 돌연변이로 진단된 Apert 증후군 1례

이영진(Young Jin Lee),고정민(Jung Min Ko),박성식(Seong Shik Park),전종근(Chong Kun Cheon) 대한의학유전학회 2010 대한의학유전학회지 Vol.7 No.2

Apert 증후군은 두부와 손발의 골 및 연부조직의 성장장애로 인해 발생하는 선천적 질환으로 두개골의 기형과 사지의 대칭적 합지증을 특징으로 하는 드문 질환이다. Apert 증후군은 변이된 FGFR2 유전자에 의해 발생한다고 알려져 있으며 세계적으로 S252W 돌연변이가 가장 흔하고, P253R 돌연변이는 드물게 보고되고 있다. 국내에서는 엑손 IIIa에서 S252W 돌연변이가 보고된 경우가 있다. 본 증례에서는 두개 봉합선의 조기융합으로 인한 두부의 특징적인 기형, 손과 발의 심한 합지증을 동반하는 전형적인 Apert 증후군 영아에서 FGFR2 유전자의 8번째 엑손부위에서의 P253R 돌연변이가 확인되었기에 보고하는 바이다. 향후 유전자-표현자형의 대한 연구, 발생 기전 및 치료와 관련한 분자 생물학적 연구가 더 필요할 것으로 사료된다. Apert syndrome is a rare congenital anomaly characterized by craniofacial malformations and severe symmetrical syndactyly of fingers and toes. This syndrome is caused by a genetic mutation; the S253 mutation is common, though the P253R mutation is not as frequent. Common symptoms include skeletal malformations, poor joint mobility, eye and ear problems, cleft palate, and orthodontic and other dental problems. We report a case of an infant with the common morphological features of Apert syndrome. Interestingly, she was found to have the P253R mutation in FGFR2 exon Ⅷ, which has been less commonly observed in Korea. A brief review of the literature is included.

Novel 5,6-disubstituted pyrrolo[2,3-<i>d</i>]pyrimidine derivatives as broad spectrum antiproliferative agents: Synthesis, cell based assays, kinase profile and molecular docking study

Lee, Ju-Hyeon,El-Damasy, Ashraf K.,Seo, Seon Hee,Gadhe, Changdev G.,Pae, Ae Nim,Jeong, Nakcheol,Hong, Soon-Sun,Keum, Gyochang Elsevier 2018 Bioorganic & medicinal chemistry Vol.26 No.21

<P><B>Abstract</B></P> <P>Two new series of 5-subtituted and 5,6-disubstituted pyrrolo[2,3-<I>d</I>]pyrimidine octamides (<B>4a</B>–<B>o</B> and <B>6a</B>–<B>g</B>) and their corresponding free amines <B>5a</B>–<B>m</B> and <B>7a</B>–<B>g</B> have been synthesized and biologically evaluated for their antiproliferative activity against three human cancer cell lines. The 5,6-disubstituted octamides <B>6d</B>–<B>g</B> as well as the amine derivative <B>7b</B> have shown the best anticancer activity with single digit micromolar GI<SUB>50</SUB> values over the tested cancer cells, and low cytotoxic effects (GI<SUB>50</SUB> > 10.0 µM) against HFF-1 normal cell. A structure activity relationship (SAR) study has been established and disclosed that terminal octamide moiety at C2 as well as disubstitution with fluorobenzyl piperazines at C5 and C6 of pyrrolo[2,3-<I>d</I>]pyrimidine are the key structural features prerequisite for best antiproliferative activity. Moreover, the most active member <B>6f</B> was tested for its antiproliferative activity over a panel of 60 cancer cell lines at NCI, and exhibited distinct broad spectrum anticancer activity with submicromolar GI<SUB>50</SUB> and TGI values over multiple cancer cells. Kinase profile of compound <B>6f</B> over 53 oncogenic kinases at 10 µM concentration showed its highly selective inhibitory activity towards FGFR4, Tie2 and TrkA kinases. The observed activity of <B>6f</B> against TrkA (IC<SUB>50</SUB> = 2.25 µM), FGFR4 (IC<SUB>50</SUB> = 6.71 µM) and Tie2 (IC<SUB>50</SUB> = 6.84 µM) was explained by molecular docking study, which also proposed that <B>6f</B> may be a type III kinase inhibitor, binding to an allosteric site rather than kinase hinge region. Overall, compound <B>6f</B> may serve as a promising anticancer lead compound that could be further optimized for development of potent anticancer agents.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Synthesis and <I>in vitro</I> antiproliferative activities of new pyrrolo[2,3-<I>d</I>]pyrimidines are reported. </LI> <LI> Compounds <B>6d–g</B> exerted single digit micromolar GI<SUB>50</SUB> values over the tested cancer cell lines. </LI> <LI> Compound <B>6f</B> has selective inhibitory activity towards FGFR4, Tie2 and TrkA kinases. </LI> <LI> Compound <B>6f</B> may be a type III allosteric kinase inhibitor. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Global Expanded Access Program for Pemigatinib in Patients with Previously Treated Locally Advanced or Metastatic Cholangiocarcinoma and Fibroblast Growth Factor Receptor Gene Alterations

Anouk Lindley,Gerald Prager,Michael Bitzer,Timothy C. Burn,Christine F. Lihou,Elisabeth Croft 대한암학회 2024 Cancer Research and Treatment Vol.56 No.3

Purpose Pemigatinib is a fibroblast growth factor receptor-2 (FGFR2) inhibitor approved for use in patients with previously treated cholangiocarcinoma (CCA) and FGFR2 fusions or rearrangements. This ongoing global Expanded Access Program (EAP) allows physicians in regions where pemigatinib is not commercially available to request pemigatinib for patients with locally advanced or metastatic CCA who, in the physician’s opinion, could benefit from pemigatinib treatment. Materials and Methods Eighty-nine patients from Europe, North America, and Israel were treated from January 2020 through September 2021. Results Patients had FGFR gene fusions (68.5%), rearrangements (12.4%), translocations (5.6%), amplifications (2.2%), and other alterations (11.2%). Median duration of treatment in the EAP was 4.0 months (range, 0.1 to 13.6 months). The most frequently reported adverse event (AE) was hyperphosphatemia (22.5%); the most common serious AE was cholangitis (3.4%). Treatment discontinuation was associated with reports of AEs for seven patients (7.9%). AEs associated with pemigatinib were consistent with those observed in clinical trials. Conclusion Efficacy was not assessed in this EAP. However, some patients remained on treatment for up to a year, suggesting that they observed a benefit from treatment. Patients with CCA should undergo molecular testing to identify those who could benefit from targeted treatments such as pemigatinib.