DOI: 10.5061/dryad.pk0p2ngz7
title: Targeted inhibition of kisspeptin neurons reverses hyperandrogenemia and abnormal hyperactive LH secretion in a preclinical mouse model of polycystic ovary syndrome

description: STUDY QUESTION Do hyperactive kisspeptin neurons contribute to
 abnormally high LH secretion and downstream hyperandrogenemia in
 polycystic ovary syndrome (PCOS)-like conditions and can inhibition of
 kisspeptin neurons rescue such endocrine impairments? SUMMARY ANSWER
 Targeted inhibition of endogenous kisspeptin neuron activity in a mouse
 model of PCOS reduced the abnormally hyperactive LH pulse secretion and
 hyperandrogenemia to healthy control levels. WHAT IS KNOWN ALREADY PCOS is
 a reproductive disorder characterized by hyperandrogenemia, anovulation,
 and/or polycystic ovaries, along with a hallmark feature of abnormal LH
 hyper-pulsatility, but the mechanisms underlying the endocrine impairments
 remain unclear. A chronic letrozole (LET; aromatase inhibitor) mouse model
 recapitulates PCOS phenotypes, including polycystic ovaries, anovulation,
 high testosterone, and hyperactive LH pulses. LET PCOS-like females also
 have increased hypothalamic kisspeptin neuronal activation which may drive
 their hyperactive LH secretion and hyperandrogenemia, but this has not
 been tested. STUDY DESIGN, SIZE, DURATION Transgenic KissCRE+/hM4Di female
 mice or littermates Cre− controls were treated with placebo, or chronic
 LET (50 µg/day) to induce a PCOS-like phenotype, followed by acute (once)
 or chronic (2 weeks) clozapine-N-oxide (CNO) exposure to chemogenetically
 inhibit kisspeptin cells (n = 6 to 10 mice/group). PARTICIPANTS/MATERIALS,
 SETTING, METHODS Key endocrine measures, including in vivo LH pulse
 secretion patterns and circulating testosterone levels, were assessed
 before and after selective kisspeptin neuron inhibition and compared
 between PCOS groups and healthy controls. Alterations in body weights were
 measured and pituitary and ovarian gene expression was determined by
 qRT-PCR. MAIN RESULTS AND THE ROLE OF CHANCE Acute targeted inhibition of
 kisspeptin neurons in PCOS mice successfully lowered the abnormally
 hyperactive LH pulse secretion (P &lt; 0.05). Likewise, chronic
 selective suppression of kisspeptin neuron activity reversed the
 previously high LH and testosterone levels (P &lt; 0.05) down to
 healthy control levels and rescued reproductive gene expression
 (P &lt; 0. 05). LIMITATIONS, REASONS FOR CAUTION Ovarian morphology
 was not assessed in this study. Additionally, mouse models can offer
 mechanistic insights into neuroendocrine processes in PCOS-like conditions
 but may not perfectly mirror PCOS in women. WIDER IMPLICATIONS OF THE
 FINDINGS These data support the hypothesis that overactive kisspeptin
 neurons can drive neuroendocrine PCOS-like impairments, and this may occur
 in PCOS women. Our findings complement recent clinical investigations
 using NKB receptor antagonists to lower LH in PCOS women and suggest that
 pharmacological dose-dependent modulation of kisspeptin neuron activity
 may be a valuable future therapeutic target to clinically treat
 hyperandrogenism and lower elevated LH in PCOS women.
description: All methods and analyses are described in detail in the published
 manuscript Methods section.
description: # Targeted inhibition of kisspeptin neurons reverses hyperandrogenemia and
 abnormal hyperactive LH secretion in a preclinical mouse model of
 polycystic ovary syndrome
 [https://doi.org/10.5061/dryad.pk0p2ngz7](https://doi.org/10.5061/dryad.pk0p2ngz7) ## Description of the data and file structure The data are from a study that compared hormone levels, body and tissue weights, and mRNA levels between several groups of animals that were exposed for 5 weeks to Letrozole (LET) or placebo control and which had their kisspeptin neurons inhibited (Cre+ animals) or not (Cre- animals) by chemogenetic technology, as described in detail in the Methods section of the published manuscript.   The dataset used to generate all graphs and figures for the associated manuscript are deposited here in an .xlsx file named  "All_figures_data_for_LET_CNO_study.xlsx".  The .xlsx file comprises 13 separate worksheets named with the corresponding manuscript Figure number (e.g., Fig 2A, Fig 3, Fig 4A, etc.) which indicates which specific figure or figure panels the data was used in the published manuscript, along with a short identifying text in the worksheet name (e.g. LH pulse profile, BW over time, Testosterone) which gives additional context about what the data listed in that worksheet corresponds to. Figures 1 and 7 were cartoon schematics and did not contain any data; therefore the .xlsx data file has worksheets for just Figures 2 through 6. ### Files and variables #### File: All\_figures\_data\_for\_LET\_CNO\_study.xlsx **Description:** The dataset used to generate all graphs in the figures of the associated manuscript are deposited here in a single .xlsx file named "All_figures_data_for_LET_CNO_study.xlsx". The xlsx file contains all data in the graphs presented in Figures 2 through 6, including data for blood LH levels (ng/ml), body weight (g), blood Testosterone levels (ng/dl), blood FSH levels (ng/ml), pituitary Lhb and Fshb levels, ovary weights (mg), ovarian Cyp17, Fshr, and Amh levels in three or four of following groups of female mice, depending on the experiment:  LET Cre-, LET Cre+, placebo Cre-, and placebo Cre+, all at either 0, 3, or 5 weeks of LET or placebo exposure. The .xlsx file comprises 13 separate worksheets named with the corresponding manuscript Figure number (e.g., Fig 2A, Fig 3, Fig 4A, etc.) which indicates which specific figure or figure panels that data was used for in the published manuscript, along with a short identifying text in the worksheet name (e.g. LH pulse profile, BW over time, Testosterone) which gives additional context about what the data in that worksheet corresponds to. Missing values are denoted as blank cells. ##### Variables and Abbreviations * LH, luteinizing hormone (ng/ml) * FSH, follicle stimulating hormone (ng/ml) * T, Testosterone (ng/dl) * BW, body weight (g) * ovary weight (mg) * pituitary Lhb and Fshb mRNA levels (fold change) * ovarian Cyp17, Fshr, and Amh mRNA levels (fold change) * LET, letrozole treated * Placebo, placebo treated (controls) * Cre+, Kiss1Cre+/DIflox/flox genotype * Cre-, Kiss1Cre-/DIflox/flox genotype (controls)