- Email: [email protected]
Endometriosis is a common denominator in unexplained pregnancy loss and infertility based on BCL6 testing
95% CI 1.05-15.2, p<0.05) and acne (OR 43.0; 95% CI 2.26-817.1, p<0.05) compared to AA. AA were less likely to believe that birth control would prevent further UIP than CA (OR 16.0; 95% CI 2.84 to 90, p<0.01). Astoundingly, despite their recent UIP, 13% of females in our sample stated they were not likely to use any form of contraception post-partum, which was similar across ethnic groups. CONCLUSIONS: Our preliminary data shows a disproportionate lack of positive perceptions regarding contraception among AA, which may be a factor leading to its non-use and subsequent UIP in this population. Understanding the disbelief among AAwomen that birth control effectively prevents pregnancy is the first step to the development of culturally targeted education and intervention for pre- and post-partum contraception in order to reduce UIP. Supported by: No financial support was necessary for the completion of this study. O-215 Wednesday, October 21, 2015 12:15 PM DISPARITIES IN ONCOFERTILITY KNOWLEDGE AMONG CANADIAN BREAST CANCER SURGEONS. S. Yee,a K. Glass,b S. Foong,c E. Kennedy,d M. Seminsky,e E. Warner.e aUniversity of Toronto, Toronto, ON, Canada; bCReATe Fertility Centre, Toronto, ON, Canada; cRegional Fertility Program, Calgary, AB, Canada; dMount Sinai Hospital, Toronto, ON, Canada; eSunnybrook Health Sciences Centre, Toronto, ON, Canada. OBJECTIVE: Breast cancer (BC) is the most common cancer in women of reproductive age, the majority of whom will receive gonadotoxic chemotherapy. Surgical oncologists are in a crucial position to initiate early fertility discussion to facilitate timely fertility preservation (FP) referral. However, the literature consistently shows suboptimal referral rates. Inadequate FP knowledge may be a contributor. This study examined factors associated with disparities in FP knowledge among Canadian BC surgeons. DESIGN: Prospective study, knowledge translation intervention. MATERIALS AND METHODS: SPOKE: Surgeon & Patient Oncofertility Knowledge Enhancement is one of 5 components of the panCanadian RUBY research program for young women with breast cancer (YWBC). We aim to improve breast surgeon FP knowledge and referrals. The lead BC surgeon from each of the 29 RUBY sites across Canada was invited to participate in a pre-intervention semi-structured telephone interview. Demographics, FP knowledge, attitudes & practice data were collected. Surgeon knowledge (based on 7 questions) was rated as good or inadequate by 2 independent investigators blinded to other responses. RESULTS: A total of 28/29 surgeons (97%) participated. Twenty (71%) stated that they routinely or usually discussed fertility issues, while 8 (29%) infrequently or never had discussions. Although all surgeons were aware of the negative impact of chemotherapy on fertility, 13 (46%) had inadequate FP knowledge. Among the 15 surgeons who were rated to have good FP knowledge, 80% routinely or usually discussed fertility issues with their patients versus 62% of the 13 others. Good FP knowledge was significantly associated with higher percentage of practice devoted to BC (p< .05). Strong trends (p¼0.06) were seen for an association between female gender, younger age and better knowledge (see table). CONCLUSIONS: We identified disparities in oncofertility knowledge among Canadian BC surgeons who treat YWBC which may affect FP discussions and referrals. In the next phase of the SPOKE study, surgeons will be provided with an oncofertility ‘‘toolbox’’ including a lecture, a one page FP option grid with versions for surgeon and patient, as well as individual site troubleshooting. Successful modification of knowledge and practice patterns by our interventions will be evaluated in 2-3 years.
Surgeon Demographics Female gender (n¼16) < 45 years old (n¼14) >¼ 50% of practice devoted to breast cancer (n¼19)
Good Knowledge (n¼15)
Inadequate Knowledge (n¼13)
Chi-square p-value
11 (73%) 10 (67%) 13 (87%)
5 (39%) 4 (31%) 6 (46%)
.06 .06 .02
Supported by: Canadian Breast Cancer Foundation & Canadian Institutes of Health Research (#OBW139590).
FERTILITY & STERILITYÒ
O-216 Wednesday, October 21, 2015 12:30 PM REGIONAL DIFFERENCES IN ELECTIVE SINGLE EMBRYO TRANSFER: IS LOCATION EVERYTHING?. J. D. Kapfhamer,a K. M. Summers,a G. Ryan,a E. M. Munch,a B. Collura,b G. D. Adamson.c aUniversity of Iowa Carver College of Medicine, Iowa City, IA; bRESOLVE: The National Infertility Association, McLean, VA; c PAMF Fertility Physicians of Northern California, Saratoga, CA. OBJECTIVE: Previous studies suggested significant regional differences in access to United States IVF centers(1), and some suggested that limited access is associated with patient decisions regarding number of embryos to transfer. Our objective was to investigate whether attitudes regarding elective single embryo transfer (eSET) and multifetal reduction are associated with geographic region. DESIGN: Retrospective descriptive analysis of cross-sectional on-line survey. MATERIALS AND METHODS: An anonymous survey regarding eSET preferences and experiences was distributed through social media over a fiveweek span in 2014. This study focused on survey participants who completed at least one IVF cycle with embryo(s) transferred or who were planning to undergo IVF. Participants were grouped into US census regions (West, Midwest, South, and Northeast). Outcome variables included preference for single embryo transfer (defined as transferring one embryo when multiple were available) vs. multiple embryo transfer (MET), and potential likelihood of undergoing multifetal reduction if advised by a provider. Bivariate analyses were performed using chi-squared for comparison of proportions among groups. Significant associations (p< .05) were included in a subsequent logistic regression analysis. RESULTS: 759 of 888 participants met inclusion criteria. The largest percentage of survey participants (36%) came from the South, with 26%, 21% and 17% percent from the Midwest, Northeast and West respectively. When compared to the Midwestern participants, participants in the South (OR 1.96[1.14-3.38]), West (OR 2.74[1.47-5.1]) and Northeast (OR 1.96[1.07-3.61]) were significantly more likely to express a preference for eSET. Participants in the Northeast would be more likely to consider multifetal reduction (OR 1.86[1.18-2.95]) when compared to the Midwestern participants. There were no differences between the Midwest, South, and West with regard to the question of multifetal reduction. CONCLUSIONS: Patients in the Midwest may be the least likely to elect eSET when compared with the other 3 US census regions. This may be due to sociocultural differences or to relative rurality and limited IVF access, making it more difficult for patients to travel for multiple IVF cycles. Targeted educational efforts may be particularly useful in this region. At the same time, patients in the Midwest may be less likely to consider multifetal reduction, at least in comparison to those in the Northeast. This may also be a result of sociocultural/religious differences or to a relative lack of education regarding the risks of multiples, and it represents another opportunity for targeted education in this region. References: 1. Nangia AK, Likosky DS, Wang D. Access to assisted reproductive technology centers in the United States. Fertility and Sterility 2010;93(3)745-61. Supported by: 1. Auxogyn, Inc., Menlo Park, CA. 2. RESOLVE: The National Infertility Association. EARLY PREGNANCY 2 O-217 Wednesday, October 21, 2015 11:15 AM ENDOMETRIOSIS IS A COMMON DENOMINATOR IN UNEXPLAINED PREGNANCY LOSS AND INFERTILITY BASED ON R. Crowe,a S. L. Young,b BCL6 TESTING. D. Slizewski,a B. A. Lessey.a aOb-Gyn, Greenville Health System, Greenville, SC; bObGyn, UNC School of Medicine, Chapel Hill, NC. OBJECTIVE: Endometriosis affects over 176 million women and is associated with infertility and pelvic pain. BCL6 is a transcriptional repressor that could confer progesterone resistance. Previously presented data suggest that BCL6 immunostaining can predict endometriosis with sensitivity and specificity that exceed 93%. Our objective was to examine the prevalence of endometriosis and other causes of implantation failure in unexplained early pregnancy loss and compare that to unexplained infertility and fertile controls. DESIGN: Laboratory analysis of prospectively collected, mid-luteal endometrium from normal fertile women, those with unexplained infertility (UI), and those unexplained recurrent pregnancy loss (uRPL). MATERIALS AND METHODS: Subjects were recruited under an IRB approved protocol. Each subject underwent LH-timed endometrial biopsy. BCL6 RNA and protein were examined using immunohistochemistry
e83
(evaluated by HSCORE) and real-time RT-PCR (evaluated for relative expression using the delta-delta Ct method). An optimal HSCORE threshold of 1.4 was previously established by ROC analysis. Differences in proportions of positive staining were analyzed by Chi-square, while RNA data were evaluated by ANOVA and T-test. RESULTS: Age and BMI were similar between the three groups. HSCORE was significantly different (p < 0.0001) (Table 1). Endometrial BCL6 immunostaining was positive (above the 1.4 HSCORE threshold) in 2/28 (7%) fertile control subjects as compared to 103/116 (88.7%) UI subjects and 32/43 (74.4%) uRPL subjects. Mean and median expression of BCL6 RNA was lowest in the fertile controls, intermediate in the uRPL and highest in UI, with ANOVA analysis approaching significance (p¼0.07). In simple pairwise comparison, mRNA levels in UI were 2-fold of those in controls (p¼0.04). Subsequent surgical exploration of a subset of subjects demonstrated endometriosis or hydrosalpinges in 98% and 89% in those with positive BCL6 immmunostaining and UI or uRPL, respectively. CONCLUSIONS: Endometrial immunostaining for BCL6 closely correlated with the fertility status of those tested, with fertile women demonstrating low levels of nuclear BCL6 and the majority of women with UI and uRPL expressing high levels. Endometrial BCL6 mRNA expression showed similar trends, but differences between groups were smaller. Additionally, high BCL6 immunostaining strongly predicted the presence of endometriosis or, occasionally, hydrosalpinges. Early pregnancy loss and Infertility appear to share a common pathophysiological feature, but may differ by severity of the defect.
asymptomatic parents, and if inherited, carry a 50% recurrence risk with highly variable clinical outcomes. Lastly, isolated deletions/duplications in 22 cases may be causative of the loss. Future studies are needed to analyze parental chromosome study outcomes and to address recurrence risk counseling. This study highlights that SNP-array-based POC studies allow for identification of small but clinically significant chromosome findings, which may be related to the cause of miscarriage and/or carry significant reproductive risks for a couple. Syndromic Microdeletions and Microduplications
Sydnromic Microdeletion
1p36 deletion1 3* 1q43 deletion3 1 5p15.33p15.2 deletion 3* (Cri du chat syndrome)5 8p23.1 deletion7 16p13.11 deletion9 16p13.11 deletion917p13.3 (Miller-Dieker syndrome)11 22q11.2 deletion(DiGeorge syndrome/VCFS)13 22q13.31 deletion (Phelan-McDermid syndrome)15 Xq22.2 deletion16
Demographics
Characteristic AGE (mean STD) BMI HSCORE
Normal Fertile Controls (n¼ 28)
Unexplained Infertility (n ¼ 116)
Unexplained Recurrent Pregnancy Loss (n ¼ 43)
32.78 2.6
33.04 4.2
34.65 4.7
25.6 4.7 0.43 0.69
25.23 5.5 2.8 1.1
25.23 6.6 2.4 1.4
Supported by: NIH R01 HD067721 (S.L.Y. and B.A.L.). O-218 Wednesday, October 21, 2015 11:30 AM SIZE MATTERS: SINGLE NUCLEOTIDE POLYMORPHISM (SNP) BASED CHROMOSOME ANALYSIS OF PRODUCTS OF CONCEPTION (POC) SAMPLES IDENTIFIES CLINICALLY SIGNIFICANT DELETIONS/DUPLICATIONS BELOW KARYOTYPE RESOLUTION. M. K. Maisenbacher,a K. Merrion,b S. Sigurjonsson,a K. G. Paik,b M. J. Young,a B. Pettersen.c aNatera, San Carlos, CA; bNatera, Inc., San Carlos, CA; cGenetic Counseling, Bend, OR. OBJECTIVE: Report on products of conception (POC) samples with deletions and duplications below the resolution of traditional karyotyping (< 10 Mb). These segmental abnormalities are clinically relevant based on size, location and/or correlation with a known genetic syndrome. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Review of 17,424 consecutive fresh POC samples sent to a reference lab along with maternal blood samples. Genotyping was performed using Illumina CytoSNP-12b microarray and bioinformatics. RESULTS: 2600 cases (14.9%) had maternal cell contamination, 17 cases (0.01%) had incomplete results, and 14,807 cases (85%) had fetal results. Of these, 100 cases (0.7%) had deletions/duplications less than 10 Mb without aneuploidy or uniparental disomy (UPD) of other chromosomes identified: 22 cases (22%) had a single isolated deletion or duplication (11 deletions and 11 duplications), 43 cases (43%) had an additional deleted or duplicated segment that was either >10 Mb or <10 Mb, and 35 cases (35%) had a specific microdeletion or microduplication syndrome (which are outlined in Table 1). CONCLUSIONS: SNP microarray with bioinformatics is a unique method for POC analysis that allows for detection of deletions/duplications below the resolution of standard karyotype analysis, i.e. less than 10 Mb. In this cohort, we identified 43 cases with a deletion and duplication, in which one or both of the segments fell below karyotype resolution; these findings are clinically relevant as parental balanced chromosome rearrangements may underlie these abnormalities. Furthermore, 35 cases of known syndromic microdeletions/duplications were found, some of which can be seen in mildly affected or
e84
ASRM Abstracts
Number of cases
Syndromic Microduplication
Number of cases
1 1 1
1q21.1 duplication2 8p23.1 duplication4 7q11.2 duplication (Williams-Beuren region duplication syndrome)6 15q11.2 duplication8 17q12 duplication10 22q11.2 duplication12
1 1 1*
6
Xp22.31 duplication14 2
2 1 9*
1 1
*
Cases with additional findings: 6/35 cases (2/3 1p36 deletion cases, 1/3 5p15.33p15.2 deletion cases and 2/9 22q11.2 duplication cases) References: 1. Battaglia A., In Pagon RA et al., GeneReviews [Internet]. Seattle (WA):University of Washington, Seattle; 1993-2015. 2008 Feb 01 [updated 2013 Jun 06]. 2. Rosenfeld JA et al., Proximal microdeletions and microduplications of 1q21.1 contribute to variable abnormal phenotypes. Eur J Hum Genet. 2012 Jul;20(7):754-61. 3. Petersen AK et al., Deletion 1q43 encompassing only CHRM3 in a patient with autistic disorder. Eur J Med Genet, 2013 Feb; 56(2):118-22. 4. Weber A et al., 8p23.1 duplication syndrome: narrowing of critical interval to 1.80 Mbp. Molecular Cytogenetics, 2014;7(1):94. 5. Mainardi CP, Cri du Chat syndrome. Orphanet J Rare Dis. 2006 Sep 5;1:33. 6. Berg JS et al., The 22q13.3 Deletion Syndrome (Phelan-McDermid Syndrome). Genet Med 2007 Jul;9(7):427-41. 7. Wat MJ et al., Chromosome 8p23.1 deletions as a cause of complex congenital heart defects and diaphragmatic hernia. Am J Med Genet Part A. 2009 Aug;149A(8):1661-77. 8. Bolton PF et al., The phenotypic manifestations of interstitial duplications of proximal 15q with special reference to the autistic spectrum disorders. Am J Med Genet 2001 Dec 8;105(8):675-85. 9. Nagamani SC et al., Phenotypic manifestations of copy number variation in chromosome 16p13.11. Eur J Hum Genet, 2011 Mar;19(3):280-6. 10. Nagamani SC et al., Clinical spectrum associated with recurrent genomic rearrangements in chromosome 17q12. Eur J Hum Genet 2010 Mar;18(3):278-84. 11. Dobyns WB et al., Lissencephaly. A human brain malformation associated with deletion of the LIS1 gene located at chromosome 17p13. JAMA 1993 Dec 15;270(230):2838-42. 12. Wentzel C et al., Clinical variability of the 22q11.2 duplication syndrome. Eur J Med Genet. 2008 Nov-Dec;51(6):501-10. 13. Emanuel et al. The 22q11.2 deletion syndrome. Adv Pediatr. 2001;48:39-73. 14. Faletra F et al., Does the 1.5 Mb microduplication in chromosome band Xp22.31 have a pathogenetic role? New contribution and a review of the literature. Am J Med Genet A. 2012 Feb;158A(2):461-4. 15. Phelan and McDermid, The 22q13.3 Deletion Syndrome (PhelanMcDermid Syndrome). Mol Syndromol 2012 Apr;2(3-5):186-201. 16. Esplin et al. Nine patients with Xp22.31 microduplication, cognitive deficits, seizures, and talipes anomalies. Am J Med Genet. 2014 Aug;164A(8):2097-103.
Vol. 104, No. 3, Supplement, September 2015
Surgeon Demographics Female gender (n¼16) < 45 years old (n¼14) >¼ 50% of practice devoted to breast cancer (n¼19)
Good Knowledge (n¼15)
Inadequate Knowledge (n¼13)
Chi-square p-value
11 (73%) 10 (67%) 13 (87%)
5 (39%) 4 (31%) 6 (46%)
.06 .06 .02
Supported by: Canadian Breast Cancer Foundation & Canadian Institutes of Health Research (#OBW139590).
FERTILITY & STERILITYÒ
O-216 Wednesday, October 21, 2015 12:30 PM REGIONAL DIFFERENCES IN ELECTIVE SINGLE EMBRYO TRANSFER: IS LOCATION EVERYTHING?. J. D. Kapfhamer,a K. M. Summers,a G. Ryan,a E. M. Munch,a B. Collura,b G. D. Adamson.c aUniversity of Iowa Carver College of Medicine, Iowa City, IA; bRESOLVE: The National Infertility Association, McLean, VA; c PAMF Fertility Physicians of Northern California, Saratoga, CA. OBJECTIVE: Previous studies suggested significant regional differences in access to United States IVF centers(1), and some suggested that limited access is associated with patient decisions regarding number of embryos to transfer. Our objective was to investigate whether attitudes regarding elective single embryo transfer (eSET) and multifetal reduction are associated with geographic region. DESIGN: Retrospective descriptive analysis of cross-sectional on-line survey. MATERIALS AND METHODS: An anonymous survey regarding eSET preferences and experiences was distributed through social media over a fiveweek span in 2014. This study focused on survey participants who completed at least one IVF cycle with embryo(s) transferred or who were planning to undergo IVF. Participants were grouped into US census regions (West, Midwest, South, and Northeast). Outcome variables included preference for single embryo transfer (defined as transferring one embryo when multiple were available) vs. multiple embryo transfer (MET), and potential likelihood of undergoing multifetal reduction if advised by a provider. Bivariate analyses were performed using chi-squared for comparison of proportions among groups. Significant associations (p< .05) were included in a subsequent logistic regression analysis. RESULTS: 759 of 888 participants met inclusion criteria. The largest percentage of survey participants (36%) came from the South, with 26%, 21% and 17% percent from the Midwest, Northeast and West respectively. When compared to the Midwestern participants, participants in the South (OR 1.96[1.14-3.38]), West (OR 2.74[1.47-5.1]) and Northeast (OR 1.96[1.07-3.61]) were significantly more likely to express a preference for eSET. Participants in the Northeast would be more likely to consider multifetal reduction (OR 1.86[1.18-2.95]) when compared to the Midwestern participants. There were no differences between the Midwest, South, and West with regard to the question of multifetal reduction. CONCLUSIONS: Patients in the Midwest may be the least likely to elect eSET when compared with the other 3 US census regions. This may be due to sociocultural differences or to relative rurality and limited IVF access, making it more difficult for patients to travel for multiple IVF cycles. Targeted educational efforts may be particularly useful in this region. At the same time, patients in the Midwest may be less likely to consider multifetal reduction, at least in comparison to those in the Northeast. This may also be a result of sociocultural/religious differences or to a relative lack of education regarding the risks of multiples, and it represents another opportunity for targeted education in this region. References: 1. Nangia AK, Likosky DS, Wang D. Access to assisted reproductive technology centers in the United States. Fertility and Sterility 2010;93(3)745-61. Supported by: 1. Auxogyn, Inc., Menlo Park, CA. 2. RESOLVE: The National Infertility Association. EARLY PREGNANCY 2 O-217 Wednesday, October 21, 2015 11:15 AM ENDOMETRIOSIS IS A COMMON DENOMINATOR IN UNEXPLAINED PREGNANCY LOSS AND INFERTILITY BASED ON R. Crowe,a S. L. Young,b BCL6 TESTING. D. Slizewski,a B. A. Lessey.a aOb-Gyn, Greenville Health System, Greenville, SC; bObGyn, UNC School of Medicine, Chapel Hill, NC. OBJECTIVE: Endometriosis affects over 176 million women and is associated with infertility and pelvic pain. BCL6 is a transcriptional repressor that could confer progesterone resistance. Previously presented data suggest that BCL6 immunostaining can predict endometriosis with sensitivity and specificity that exceed 93%. Our objective was to examine the prevalence of endometriosis and other causes of implantation failure in unexplained early pregnancy loss and compare that to unexplained infertility and fertile controls. DESIGN: Laboratory analysis of prospectively collected, mid-luteal endometrium from normal fertile women, those with unexplained infertility (UI), and those unexplained recurrent pregnancy loss (uRPL). MATERIALS AND METHODS: Subjects were recruited under an IRB approved protocol. Each subject underwent LH-timed endometrial biopsy. BCL6 RNA and protein were examined using immunohistochemistry
e83
(evaluated by HSCORE) and real-time RT-PCR (evaluated for relative expression using the delta-delta Ct method). An optimal HSCORE threshold of 1.4 was previously established by ROC analysis. Differences in proportions of positive staining were analyzed by Chi-square, while RNA data were evaluated by ANOVA and T-test. RESULTS: Age and BMI were similar between the three groups. HSCORE was significantly different (p < 0.0001) (Table 1). Endometrial BCL6 immunostaining was positive (above the 1.4 HSCORE threshold) in 2/28 (7%) fertile control subjects as compared to 103/116 (88.7%) UI subjects and 32/43 (74.4%) uRPL subjects. Mean and median expression of BCL6 RNA was lowest in the fertile controls, intermediate in the uRPL and highest in UI, with ANOVA analysis approaching significance (p¼0.07). In simple pairwise comparison, mRNA levels in UI were 2-fold of those in controls (p¼0.04). Subsequent surgical exploration of a subset of subjects demonstrated endometriosis or hydrosalpinges in 98% and 89% in those with positive BCL6 immmunostaining and UI or uRPL, respectively. CONCLUSIONS: Endometrial immunostaining for BCL6 closely correlated with the fertility status of those tested, with fertile women demonstrating low levels of nuclear BCL6 and the majority of women with UI and uRPL expressing high levels. Endometrial BCL6 mRNA expression showed similar trends, but differences between groups were smaller. Additionally, high BCL6 immunostaining strongly predicted the presence of endometriosis or, occasionally, hydrosalpinges. Early pregnancy loss and Infertility appear to share a common pathophysiological feature, but may differ by severity of the defect.
asymptomatic parents, and if inherited, carry a 50% recurrence risk with highly variable clinical outcomes. Lastly, isolated deletions/duplications in 22 cases may be causative of the loss. Future studies are needed to analyze parental chromosome study outcomes and to address recurrence risk counseling. This study highlights that SNP-array-based POC studies allow for identification of small but clinically significant chromosome findings, which may be related to the cause of miscarriage and/or carry significant reproductive risks for a couple. Syndromic Microdeletions and Microduplications
Sydnromic Microdeletion
1p36 deletion1 3* 1q43 deletion3 1 5p15.33p15.2 deletion 3* (Cri du chat syndrome)5 8p23.1 deletion7 16p13.11 deletion9 16p13.11 deletion917p13.3 (Miller-Dieker syndrome)11 22q11.2 deletion(DiGeorge syndrome/VCFS)13 22q13.31 deletion (Phelan-McDermid syndrome)15 Xq22.2 deletion16
Demographics
Characteristic AGE (mean STD) BMI HSCORE
Normal Fertile Controls (n¼ 28)
Unexplained Infertility (n ¼ 116)
Unexplained Recurrent Pregnancy Loss (n ¼ 43)
32.78 2.6
33.04 4.2
34.65 4.7
25.6 4.7 0.43 0.69
25.23 5.5 2.8 1.1
25.23 6.6 2.4 1.4
Supported by: NIH R01 HD067721 (S.L.Y. and B.A.L.). O-218 Wednesday, October 21, 2015 11:30 AM SIZE MATTERS: SINGLE NUCLEOTIDE POLYMORPHISM (SNP) BASED CHROMOSOME ANALYSIS OF PRODUCTS OF CONCEPTION (POC) SAMPLES IDENTIFIES CLINICALLY SIGNIFICANT DELETIONS/DUPLICATIONS BELOW KARYOTYPE RESOLUTION. M. K. Maisenbacher,a K. Merrion,b S. Sigurjonsson,a K. G. Paik,b M. J. Young,a B. Pettersen.c aNatera, San Carlos, CA; bNatera, Inc., San Carlos, CA; cGenetic Counseling, Bend, OR. OBJECTIVE: Report on products of conception (POC) samples with deletions and duplications below the resolution of traditional karyotyping (< 10 Mb). These segmental abnormalities are clinically relevant based on size, location and/or correlation with a known genetic syndrome. DESIGN: Retrospective analysis. MATERIALS AND METHODS: Review of 17,424 consecutive fresh POC samples sent to a reference lab along with maternal blood samples. Genotyping was performed using Illumina CytoSNP-12b microarray and bioinformatics. RESULTS: 2600 cases (14.9%) had maternal cell contamination, 17 cases (0.01%) had incomplete results, and 14,807 cases (85%) had fetal results. Of these, 100 cases (0.7%) had deletions/duplications less than 10 Mb without aneuploidy or uniparental disomy (UPD) of other chromosomes identified: 22 cases (22%) had a single isolated deletion or duplication (11 deletions and 11 duplications), 43 cases (43%) had an additional deleted or duplicated segment that was either >10 Mb or <10 Mb, and 35 cases (35%) had a specific microdeletion or microduplication syndrome (which are outlined in Table 1). CONCLUSIONS: SNP microarray with bioinformatics is a unique method for POC analysis that allows for detection of deletions/duplications below the resolution of standard karyotype analysis, i.e. less than 10 Mb. In this cohort, we identified 43 cases with a deletion and duplication, in which one or both of the segments fell below karyotype resolution; these findings are clinically relevant as parental balanced chromosome rearrangements may underlie these abnormalities. Furthermore, 35 cases of known syndromic microdeletions/duplications were found, some of which can be seen in mildly affected or
e84
ASRM Abstracts
Number of cases
Syndromic Microduplication
Number of cases
1 1 1
1q21.1 duplication2 8p23.1 duplication4 7q11.2 duplication (Williams-Beuren region duplication syndrome)6 15q11.2 duplication8 17q12 duplication10 22q11.2 duplication12
1 1 1*
6
Xp22.31 duplication14 2
2 1 9*
1 1
*
Cases with additional findings: 6/35 cases (2/3 1p36 deletion cases, 1/3 5p15.33p15.2 deletion cases and 2/9 22q11.2 duplication cases) References: 1. Battaglia A., In Pagon RA et al., GeneReviews [Internet]. Seattle (WA):University of Washington, Seattle; 1993-2015. 2008 Feb 01 [updated 2013 Jun 06]. 2. Rosenfeld JA et al., Proximal microdeletions and microduplications of 1q21.1 contribute to variable abnormal phenotypes. Eur J Hum Genet. 2012 Jul;20(7):754-61. 3. Petersen AK et al., Deletion 1q43 encompassing only CHRM3 in a patient with autistic disorder. Eur J Med Genet, 2013 Feb; 56(2):118-22. 4. Weber A et al., 8p23.1 duplication syndrome: narrowing of critical interval to 1.80 Mbp. Molecular Cytogenetics, 2014;7(1):94. 5. Mainardi CP, Cri du Chat syndrome. Orphanet J Rare Dis. 2006 Sep 5;1:33. 6. Berg JS et al., The 22q13.3 Deletion Syndrome (Phelan-McDermid Syndrome). Genet Med 2007 Jul;9(7):427-41. 7. Wat MJ et al., Chromosome 8p23.1 deletions as a cause of complex congenital heart defects and diaphragmatic hernia. Am J Med Genet Part A. 2009 Aug;149A(8):1661-77. 8. Bolton PF et al., The phenotypic manifestations of interstitial duplications of proximal 15q with special reference to the autistic spectrum disorders. Am J Med Genet 2001 Dec 8;105(8):675-85. 9. Nagamani SC et al., Phenotypic manifestations of copy number variation in chromosome 16p13.11. Eur J Hum Genet, 2011 Mar;19(3):280-6. 10. Nagamani SC et al., Clinical spectrum associated with recurrent genomic rearrangements in chromosome 17q12. Eur J Hum Genet 2010 Mar;18(3):278-84. 11. Dobyns WB et al., Lissencephaly. A human brain malformation associated with deletion of the LIS1 gene located at chromosome 17p13. JAMA 1993 Dec 15;270(230):2838-42. 12. Wentzel C et al., Clinical variability of the 22q11.2 duplication syndrome. Eur J Med Genet. 2008 Nov-Dec;51(6):501-10. 13. Emanuel et al. The 22q11.2 deletion syndrome. Adv Pediatr. 2001;48:39-73. 14. Faletra F et al., Does the 1.5 Mb microduplication in chromosome band Xp22.31 have a pathogenetic role? New contribution and a review of the literature. Am J Med Genet A. 2012 Feb;158A(2):461-4. 15. Phelan and McDermid, The 22q13.3 Deletion Syndrome (PhelanMcDermid Syndrome). Mol Syndromol 2012 Apr;2(3-5):186-201. 16. Esplin et al. Nine patients with Xp22.31 microduplication, cognitive deficits, seizures, and talipes anomalies. Am J Med Genet. 2014 Aug;164A(8):2097-103.
Vol. 104, No. 3, Supplement, September 2015